Scholarly Literature
This is an aggregation of scholarly literature from peer-reviewed journals, bioRxiv , medRxiv and arXiv preprint servers focused on natural and engineered selfish genetic elements and genetic biocontrol. New citations are added as they appear. This content is intended for anyone interested in the scholarly literature on gene drive and genetic biocontrol.
Intra-lineage microevolution of Wolbachia leads to the emergence of new cytoplasmic incompatibility patterns
28889Namias A, Ngaku A, Makoundou P, Unal S, Sicard M, Weill M, PLoS Biology, 2024-02-13 20:25:15.
Mosquitoes of the Culex pipiens complex are worldwide vectors of arbovirus, filarial nematodes, and avian malaria agents. In these hosts, the endosymbiotic bacteria Wolbachia induce cytoplasmic incompatibility (CI), i.e., reduced embryo viability in so-called incompatible crosses. Wolbachia infecting Culex pipiens (wPip) cause CI patterns of unparalleled complexity, associated with the amplification and diversification of cidA and cidB genes, with up to 6 different gene copies described in a single wPip genome. In wPip, CI is thought to function as a toxin-antidote (TA) system where compatibility relies on having the right antidotes (CidA) in the female to bind and neutralize the male’s toxins (CidB). By repeating crosses between Culex isofemale lines over a 17 years period, we documented the emergence of a new compatibility type in real time and linked it to a change in cid genes genotype. We showed that loss of specific cidA gene copies in some wPip genomes results in a loss of compatibility. More precisely, we found that this lost antidote had an original sequence at its binding interface, corresponding to the original sequence at the toxin’s binding interface. We showed that these original cid variants are recombinant, supporting a role for recombination rather than point mutations in rapid CI evolution. These results strongly support the TA model in natura, adding to all previous data acquired with transgenes expression.
Transgenic expression of cif genes from Wolbachia strain wAlbB recapitulates cytoplasmic incompatibility in Aedes aegypti
28882McNamara, C.J., Ant, T.H., Harvey-Samuel, T. et al., Nature Communications, 15. 2024-02-13 17:43:47.
The endosymbiotic bacteria Wolbachia can invade insect populations by modifying host reproduction through cytoplasmic incompatibility (CI), an effect that results in embryonic lethality when Wolbachia-carrying males mate with Wolbachia-free females. Here we describe a transgenic system for recreating CI in the major arbovirus vector Aedes aegypti using CI factor (cif) genes from wAlbB, a Wolbachia strain currently being deployed to reduce dengue transmission. CI-like sterility is induced when cifA and cifB are co-expressed in testes; this sterility is rescued by maternal cifA expression, thereby reproducing the pattern of Wolbachia-induced CI. Expression of cifB alone is associated with extensive DNA damage and disrupted spermatogenesis. The strength of rescue by maternal cifA expression is dependent on the comparative levels of cifA/cifB expression in males. These findings are consistent with CifB acting as a toxin and CifA as an antitoxin, with CifA attenuating CifB toxicity in both the male germline and in developing embryos. These findings provide important insights into the interactions between cif genes and their mechanism of activity and provide a foundation for the building of a cif gene-based drive system in Ae. aegypti.
Transforming malaria prevention and control: the prospects and challenges of gene drive technology for mosquito management
28861Yusuf Amuda Tajudeen, Habeebullah Jayeola Oladipo, Iyiola Olatunji Oladunjoye, Muhammad Kamaldeen Oladipo, Hameedat Damilola Shittu, Imam-Fulani Abdulmumeen, Abdullateef Opeyemi Afolabi and Mona Said El-Sherbini, Annals of Medicine, 55. 2024-01-30 17:32:22.
In the era of insecticides and anti-malarial drug resistance, gene drive technology holds considerable promise for malaria control. Gene drive technology deploys genetic modifications into mosquito populations to impede their ability to transmit the malaria parasite. This can be either through the disruption of an essential mosquito gene or the association of gene drive with a desirable effector gene. CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is a gene editing tool that precisely modifies mosquito vector DNA sequences and curtails the rate of pathogen transmission. A comprehensive search was conducted in the SCOPUS and MEDLINE databases (via PubMed) until October 2023. The keywords used were related to the principles and mechanisms of gene drive technology, its advantages, and disadvantages, and its ethical and regulatory considerations in sustainable malaria eradication. The development of gene drive enables the preferential inheritance of specific genes in targeted mosquitoes, potentially obstructing the transmission of the Plasmodium parasite. This technology was also studied for the control of other vector-borne diseases such as dengue and chikungunya viruses. Despite its experimental superiority over other traditional methods such as insecticide-treated nets and insecticide sprays, the long-term dynamic interplay of mutation and resistance poses challenges for gene drive efficiency in sustainable malaria control. This commentary elucidates the underlying mechanisms and principles of gene drive technology, underscoring its promise and challenges as a novel strategy to curtail malaria prevalence. Although the release of such genetically modified mosquitoes into the natural environment would result in the eradication of the locally targeted species of mosquitoes, the complete eradication of the entire species remains questionable. Thus, the practical application raises significant ethical and regulatory concerns for further research and risk assessment, including the risk of gene drive spreading to nontarget species in the wider theatre of biodiverse species.
A multiplexed, confinable CRISPR/Cas9 gene drive can propagate in caged Aedes aegypti populations
28857Anderson, M.A.E., Gonzalez, E., Edgington, M.P. et al., Nature Communications, 15. 2024-01-30 16:38:51.
Aedes aegypti is the main vector of several major pathogens including dengue, Zika and chikungunya viruses. Classical mosquito control strategies utilizing insecticides are threatened by rising resistance. This has stimulated interest in new genetic systems such as gene drivesHere, we test the regulatory sequences from the Ae. aegypti benign gonial cell neoplasm (bgcn) homolog to express Cas9 and a separate multiplexing sgRNA-expressing cassette inserted into the Ae. aegypti kynurenine 3-monooxygenase (kmo) gene. When combined, these two elements provide highly effective germline cutting at the kmo locus and act as a gene drive. Our target genetic element drives through a cage trial population such that carrier frequency of the element increases from 50% to up to 89% of the population despite significant fitness costs to kmo insertions. Deep sequencing suggests that the multiplexing design could mitigate resistance allele formation in our gene drive system.
Evidence of Differences in Cellular Regulation of Wolbachia-Mediated Viral Inhibition between Alphaviruses and Flaviviruses
28852Rainey SM, Lefteri DA, Darby C, Kohl A, Merits A, Sinkins SP., Viruses, 16(1):115. 2024-01-28 13:51:52.
The intracellular bacterium Wolbachia is increasingly being utilised in control programs to limit the spread of arboviruses by Aedes mosquitoes. Achieving a better understanding of how Wolbachia strains can reduce viral replication/spread could be important for the long-term success of such programs. Previous studies have indicated that for some strains of Wolbachia, perturbations in lipid metabolism and cholesterol storage are vital in Wolbachia-mediated antiviral activity against the flaviviruses dengue and Zika; however, it has not yet been examined whether arboviruses in the alphavirus group are affected in the same way. Here, using the reporters for the alphavirus Semliki Forest virus (SFV) in Aedes albopictus cells, we found that Wolbachia strains wMel, wAu and wAlbB blocked viral replication/translation early in infection and that storage of cholesterol in lipid droplets is not key to this inhibition. Another alphavirus, o’nyong nyong virus (ONNV), was tested in both Aedes albopictus cells and in vivo in stable, transinfected Aedes aegypti mosquito lines. The strains wMel, wAu and wAlbB show strong antiviral activity against ONNV both in vitro and in vivo. Again, 2-hydroxypropyl-β-cyclodextrin (2HPCD) was not able to rescue ONNV replication in cell lines, suggesting that the release of stored cholesterol caused by wMel is not able to rescue blockage of ONNV. Taken together, this study shows that alphaviruses appear to be inhibited early in replication/translation and that there may be differences in how alphaviruses are inhibited by Wolbachia in comparison to flaviviruses.
Stakeholder engagement in the development of genetically modified mosquitoes for malaria control in West Africa: lessons learned from 10 years of Target Malaria’s work in Mali
28850Bakara Dicko, Souleymane Kodio, Hatouma Samoura, Fatoumata Traoré, Naima Sykes, Mouhamed Drabo, Delphine Thizy, Isabelle Coche, Benjamin Robinson, Kadiatou Sanogo, Bilkissou Yagouré, Samba Diop, Mamadou B. Coulibaly, Frontiers in Bioengineering and Biotechnology, 11. 2024-01-28 13:28:47.
From 2012 to 2023, the Malaria Research and Training Center (MRTC), based out of the University of Sciences, Techniques and Technologies of Bamako (USTTB), was part of the Target Malaria research consortium working towards developing novel gene drive-based tools for controlling populations of malaria vector mosquitoes. As part of this work, Target Malaria Mali has undertaken a range of in-depth engagement activities with the communities where their research is conducted and with other stakeholders nationally. These activities were meant to ensure that the project’s activities took place with the agreement of those communities, and that those communities were able to play a role in shaping the project’s approach to ensure that its eventual outcomes were in line with their needs and concerns. This paper aims to conduct a critical assessment of those 10 years of stakeholder engagement in order to identify good practices which can inform future engagement work on gene drive research in West Africa. It sets out a range of approaches and practices that enabled the Target Malaria Mali team to engage a variety of stakeholders, to share information, collect feedback, and determine community agreement, in a manner that was inclusive, effective, and culturally appropriate. These can be useful tools for those working on gene drive research and other area-wide vector control methods in West African contexts to ensure that their research is aligned with the interests of the communities who are intended to be its ultimate beneficiaries, and to allow those communities to play a meaningful role in the research process.
The haplolethal gene wupA of Drosophila exhibits potential as a target for an X-poisoning gene drive
28845Clancy D. Lawler, Ana Karla Parra Nuñez, Natalia Hernandes, Soumitra Bhide, Isabelle Lohrey, Simon Baxter, Charles Robin, bioRxiv, 2024-01-28 12:54:44.
A synthetic gene drive that targets haplolethal genes on the X-chromosome can skew the sex ratio towards males. Like an ‘X-shredder’ it does not involve ‘homing’ and that has advantages including the reduction of gene drive resistance allele formation. We examine this ‘X-poisoning’ strategy by targeting four of the 11 known X-linked haplolethal/haplosterile genes of Drosophila melanogaster with CRISPR/Cas9. We find that targeting the wupA gene during spermatogenesis skews the sex ratio so fewer than 14% of progeny are daughters. That is unless we cross the mutagenic males to X^XY female flies that bear attached-X chromosomes, which reverses the inheritance of the poisoned X chromosome so that sons inherit it from their father; in which case only 2% of the progeny are sons. These sex ratio biases suggests that most of the CRISPR/Cas9 mutants we induced in the wupA gene are haplolethal but some are recessive lethal. The males generating wupA mutants do not suffer from reduced fertility rather the haplolethal mutants arrest development in the late stages of embryogenesis well after fertilized eggs have been laid. This provides a distinct advantage over genetic manipulation strategies involving sterility which can be countered by the remating of females. We also find that wupA mutants that destroy the nuclear localization signal of shorter isoforms are not haplolethal as long as the open reading frame remains intact. Like D. melanogaster wupA orthologs of D. suzukii and Anopheles mosquitos are found on X chromosomes making wupA a viable X-poisoning target in multiple species.
A bumpy road ahead for genetic biocontainment
28833George, D.R., Danciu, M., Davenport, P.W. et al, Nature Communications, 2024-01-23 18:15:26.
The environmental release of bioengineered organisms is increasingly being suggested for a variety of applications, including bioremediation, biosequestration, bio-mining, environmental biosensing and conservation. The objectives of many environmental release applications shift the goals of biocontainment from preventing organism spread outside of closed spaces (e.g., laboratories or bioreactors) to managing the persistence of engineered organisms and their genetic material in open, dynamic environments. In the scientific literature, discussions of environmental release are often accompanied by calls for robust “intrinsic biocontainment”, where containment mechanisms are genetically engineered into the organism to limit and control its spread and persistence. A number of intrinsic biocontainment approaches have so far been proposed, and can be grouped into two overarching strategies. First, gene-flow barriers attempt to limit the spread of genetic material through lateral gene transfer, which refers to the ability of cells to directly exchange DNA molecules with one another or absorb DNA from external environmental sources. This can be accomplished through conditional lethality strategies (such as toxin anti-toxin systems or targeted DNA degradation), or through limiting plasmid replication or deleting natural competence genes from target cells. Second, strain/host control strategies seek to prevent survival and growth of engineered microbes outside specific environmental conditions using growth restriction and fitness control strategies such as metabolic auxotrophy, kill switches, and conditional essentiality. Over the past decade, researchers and developers have expanded the technical toolkit of intrinsic biocontainment techniques, with new approaches using orthogonal sequences, synthetic auxotrophy, CRISPR-based kill switches, sequence-entanglement, and “cell-free” systems.
The epidemiology of imported and locally-acquired dengue in Australia, 2012–2022
28826Asma Sohail, Katherine L Anders, Sarah L McGuinness, Karin Leder, Journal of Travel Medicine, 2024-01-23 17:13:52.
Dengue is the most important arboviral disease globally, and poses ongoing challenges for control including in non-endemic countries with competent mosquito vectors at risk of local transmission through imported cases. We examined recent epidemiological trends in imported and locally-acquired dengue in Australia, where the Wolbachia mosquito population replacement method was implemented throughout dengue-prone areas of northern Queensland between 2011–2019. Dengue is the most important arboviral disease globally, and poses ongoing challenges for control including in non-endemic countries with competent mosquito vectors at risk of local transmission through imported cases. We examined recent epidemiological trends in imported and locally-acquired dengue in Australia, where the Wolbachia mosquito population replacement method was implemented throughout dengue-prone areas of northern Queensland between 2011–2019. Dengue is the most important arboviral disease globally, and poses ongoing challenges for control including in non-endemic countries with competent mosquito vectors at risk of local transmission through imported cases. We examined recent epidemiological trends in imported and locally-acquired dengue in Australia, where the Wolbachia mosquito population replacement method was implemented throughout dengue-prone areas of northern Queensland between 2011–2019.
Recombinant venom proteins in insect seminal fluid reduces female lifespan
28821Samuel J. Beach, Maciej Maselko, bioRxiv, 2024-01-23 14:49:23.
The emergence of insecticide resistance has increased the need for alternative pest management tools1,2 Numerous genetic biocontrol approaches, which involve the release of genetically modified organisms to control pest populations, are in various stages of development to provide highly targeted pest control3-7. However, all current mating-based genetic biocontrol technologies function by releasing engineered males which skew sex-ratios or reduce offspring viability in subsequent generations. This allows mated females continue to cause harm (e.g. transmit disease). Here, we demonstrate the first example of intragenerational genetic biocontrol, wherein mating with engineered males reduces female lifespan. The toxic male technique (TMT) involves the heterologous expression of insecticidal proteins within the male reproductive tract that are transferred to females via mating. We demonstrate TMT in Drosophila melanogaster males, which reduce the median lifespan of mated females by 37 - 59% compared to controls mated to wild type males. Agent-based models of Aedes aegypti predict that compared to existing genetic biocontrol technologies, even modest levels of mated female mortality could allow TMT to suppress a female population substantially faster, which is likely to result in reduced disease burdens. TMT presents a novel approach for combatting outbreaks of disease vectors and agricultural pests.
Maxizyme-mediated suppression of chikungunya virus replication and transmission in transgenic Aedes aegypti mosquitoes
28819Mishra P, Balaraman V, Fraser Jr. M, Frontiers in Microbiology, 14. 2024-01-23 14:40:52.
Chikungunya virus (CHIKV) is an emerging mosquito-borne pathogen of significant public health importance. There are currently no prophylactic vaccines or therapeutics available to control CHIKV. One approach to arbovirus control that has been proposed is the replacement of transmission-competent mosquitoes with those that are refractory to virus infection. Several transgene effectors are being examined as potentially useful for this population replacement approach. We previously demonstrated the successful use of hammerhead ribozymes (hRzs) as an antiviral effector transgene to control CHIKV infection of, and transmission by, Aedes mosquitoes. In this report we examine a maxizyme approach to enhance the catalytic activity and prevent virus mutants from escaping these ribozymes. We designed a maxizyme containing minimized (monomer) versions of two hRzs we previously demonstrated to be the most effective in CHIKV suppression. Three versions of CHIKV maxizyme were designed: Active (Mz), inactive (ΔMz), and a connected CHIKV maxizyme (cMz). The maxizymes with their expression units (Ae-tRNA val promoter and its termination signal) were incorporated into lentivirus vectors with selection and visualization markers. Following transformation, selection, and single-cell sorting of Vero cells, clonal cell populations were infected with CHIKV at 0.05 and 0.5 MOI, and virus suppression was assessed using TCID50-IFA, RT-qPCR, and caspase-3 assays. Five transgenic mosquito lines expressing cMz were generated and transgene insertion sites were confirmed by splinkerette PCR. Our results demonstrate that Vero cell clones expressing Mz exhibited complete inhibition of CHIKV replication compared to their respective inactive control version or the two parent hRzs. Upon oral challenge of transgenic mosquitoes with CHIKV, three out of the five lines were completely refractory to CHIKV infection, and all five lines tested negative for salivary transmission. Altogether, this study demonstrates that maxizymes can provide a higher catalytic activity and viral suppression than hRzs.
Mass-Rearing Conditions Do Not Always Reduce Genetic Diversity: The Case of the Mexican Fruit Fly, Anastrepha ludens (Diptera: Tephritidae)
28817Ruiz-Montoya L, Sánchez-Rosario M, López-Gómez E, Garcia-Bautista M, Canedo-Texón A, Haymer D, Liedo P, Insects, 15:56. 2024-01-23 14:31:12.
The application of the sterile insect technique (SIT) requires the adaptation of insects to mass-rearing conditions. It is generally accepted that this adaptation may include a reduction in genetic diversity and an associated loss of desirable characteristics for the effective performance of sterile insects in the field. Here, we compare the genetic diversity of two mass-reared strains of the Mexican fruit fly, Anastrepha ludens, and a wild (WIL) population collected near Tapachula, Mexico, using seven DNA microsatellites as molecular genetic markers. The mass-reared strains were a bisexual laboratory strain (LAB) with approximately 130 generations under mass-rearing and a genetic sexing strain, Tapachula-7 (TA7), also under mass-rearing for 100 generations. Our results revealed an overall low level of genetic differentiation (approximately 15%) among the three strains, with the LAB and WIL populations being genetically most similar and TA7 most genetically differentiated. Although there were some differences in allele frequencies between strains, our results show that overall, the adaptation to mass-rearing conditions did not reduce genetic variability compared to the wild sample in terms of heterozygosity or allelic richness, nor did it appear to alter the level of inbreeding with respect to the wild populations. These results are contrary to the general idea that mass-rearing always results in a reduction in genetic diversity. Overall, our findings can contribute to a better understanding of the impact that adaptation to mass-rearing conditions may have on the genetic make-up of strains.
Wolbachia still works when it is warm
28809McKay, A., Nature Ecology and Evolution, 8. 2024-01-16 13:59:40.
Wolbachia is a maternally inherited endosymbiotic bacterium that can impede the transmission of viruses such as dengue and Zika by some mosquito vectors to humans. Over the past decade, this self-sustaining disease-control method has been rolled out in cities of increasing size; 2023 saw the largest demonstration of efficacy to date, in Medellín, Colombia. However, two key biological mechanisms that enable Wolbachia-carrying mosquitoes to spread in a population — cytoplasmic incompatibility and maternal transmission — have been shown in laboratory conditions to decline under high temperatures. Writing in Nature Climate Change in August 2023, Vásquez and colleagues conducted computational simulations to explore whether near-future warming scenarios are likely to affect the efficacy of Wolbachia-based biocontrol. The models combine empirical estimates of thermal sensitivity for one strain of Wolbachia with projections of future average temperatures and heatwaves for Cairns, Australia and Nha Trang City, Vietnam, two locations where the intervention has been deployed. Estimates of the bacterium’s successful persistence in the mosquito population remain high under tested projections of moderate and severe average warming to 2050. However, the simulations also show reduced efficacy under more variable thermal extremes; these interventions have the potential to fail under extreme warming with long heatwave durations. We selected this paper for our Year in Review collection because it exemplifies how insect thermal biology and global change analysis can offer insights relevant to human health. Testing whether climate change influences Wolbachia-based biocontrol helps to illuminate whether the promising technology is likely to remain feasible in coming years.
Gene drive and genetic sex conversion in the global agricultural pest Ceratitis capitata
28802Meccariello, A., Hou, S., Davydova, S. et al., Nature Communications, 15:372. 2024-01-15 17:13:20.
Homing-based gene drives are recently proposed interventions promising the area-wide, species-specific genetic control of harmful insect populations. Here we characterise a first set of gene drives in a tephritid agricultural pest species, the Mediterranean fruit fly Ceratitis capitata (medfly). Our results show that the medfly is highly amenable to homing-based gene drive strategies. By targeting the medfly transformer gene, we also demonstrate how CRISPR-Cas9 gene drive can be coupled to sex conversion, whereby genetic females are transformed into fertile and harmless XX males. Given this unique malleability of sex determination, we modelled gene drive interventions that couple sex conversion and female sterility and found that such approaches could be effective and tolerant of resistant allele selection in the target population. Our results open the door for developing gene drive strains for the population suppression of the medfly and related tephritid pests by co-targeting female reproduction and shifting the reproductive sex ratio towards males. They demonstrate the untapped potential for gene drives to tackle agricultural pests in an environmentally friendly and economical way.
Programmed evolution: Using asexual gene drives to sculpt tumor populations and combat genetic diversity
28800Justin Pritchard, Scott Leighow, Joshua Reynolds et al., Research Square, 2024-01-15 17:07:16.
Resistance evolution is responsible for the failure of most targeted anticancer therapies. Tumor heterogeneity is so profound that pre-existing resistance is thought to be guaranteed at the time that advanced disease is detected. The practice of waiting for treatment failure and then responding to the refractory tumor with next-generation targeted therapies locks clinicians in an evolutionary arms race until no further treatment options are available. Here, we posit that disease evolution can be reproducibly reprogrammed to design more readily treated tumors, regardless of the exact ensemble of pre-existing genetic heterogeneity. To this end, we conceive of a modular genetic platform that couples an inducible fitness advantage with a shared fitness cost. Using stochastic models of evolutionary dynamics, we identify the design criteria of these “selection gene drives.” We then build prototypes that leverage the selective pressure of various approved tyrosine kinase inhibitors and employ therapeutic mechanisms as diverse as prodrug catalysis and immune activity induction. By using saturating mutagenesis across a drug target and genome-scale loss-of-function libraries, we show that our selection gene drives can eradicate extremely diverse forms of genetic resistance. Finally, using theory to guide treatment scheduling, we demonstrate that model-informed switch engagement can create dramatic in vivo efficacy. These results establish selection gene drives as a powerful new paradigm for evolution-guided anticancer therapy.
Sex or poison? Genetic pest management in the 21st century
28782Luke Alphey, BMC Biology, 21:289. 2024-01-11 13:14:20.
Pests do enormous damage to human and animal health, to agriculture and to biodiversity, with mosquitoes transmitting pathogens, insect larvae eating crops or invasive rodents threatening the last island refuges of endangered birds. This commentary focuses on insects, particularly mosquitoes. However, most considerations apply equally to other pest species. Genetic pest management (GPM) is the use of genetics to control pests through mating of modified pests with their wildtype counterparts. This allows heritable traits to be transferred (“introgressed”) into the wild pest population. In principle, any sexually reproducing pest species can be targeted. The aim is to reduce harm done by the pest population, with typical intended outcomes overwhelmingly falling into two types: population suppression and population modification. For population suppression, one would introgress fitness-reducing traits, such as lethality or sterility, leading to reduction in the numerical size of the pest populations if spread into the target population at sufficiently high frequency. Population modification aims to reduce the harm done by the pest without large changes in the numerical size of the pest population, for example by reducing the ability to transmit disease (“vector competence”) of modified mosquitoes. If such traits, or the DNA sequences encoding them, can be sustained at sufficiently high allele frequency in the target population then the desired harm-reduction outcome should be achieved, by reduction in the number of pests or by reduction in the per-pest harm.
Limited association between Wolbachia and Plasmodium falciparum infections in natural populations of the major malaria mosquito Anopheles moucheti
28760Théo Mouillaud, Audric Berger, Marie Buysse, Nil Rahola, Josquin Daron, Jean-Pierre Agbor, Sandrine N. Sango, Daniel E. Neafsey, Olivier Duron, Diego Ayala, Evolutionary Applications, 16:1999-2006. 2024-01-07 16:47:29.
Since the discovery of natural malaria vector populations infected by the endosymbiont bacterium Wolbachia, a renewed interest has arisen for using this bacterium as an alternative for malaria control. Among naturally infected mosquitoes, Anopheles moucheti, a major malaria mosquito in Central Africa, exhibits one of the highest prevalences of Wolbachia infection. To better understand whether this maternally inherited bacterium could be used for malaria control, we investigated Wolbachia influence in An. moucheti populations naturally infected by the malaria parasite Plasmodium falciparum. To this end, we collected mosquitoes in a village from Cameroon, Central Africa, where this mosquito is the main malaria vector. We found that the prevalence of Wolbachia bacterium was almost fixed in the studied mosquito population, and was higher than previously recorded. We also quantified Wolbachia in whole mosquitoes and dissected abdomens, confirming that the bacterium is also elsewhere than in the abdomen, but at lower density. Finally, we analyzed the association of Wolbachia presence and density on P. falciparum infection. Wolbachia density was slightly higher in mosquitoes infected with the malaria parasite than in uninfected mosquitoes. However, we observed no correlation between the P. falciparum and Wolbachia densities. In conclusion, our study indicates that naturally occurring Wolbachia infection is not associated to P. falciparum development within An. moucheti mosquitoes.
The mosquito knows no borders: Regional challenges for global confrontation in the dengue battle
28789Barçante JMdP, Cherem J, PLoS Neglected Tropical Diseases, 18. 2024-01-04 14:05:29.
Dengue fever is a neglected disease with a global impact, and its incidence and geographical reach are rapidly expanding. Global warming marked by higher average temperatures, precipitation, and longer periods of drought could prompt a record number of dengue fever infections worldwide, due to the thermal biology of mosquitoes. Additionally, increased movement of people, urbanization, and pressure on water and sanitation have driven the spread of dengue fever. The absence of access to tap water leads to an increased reliance on water-storing containers, which can facilitate the breeding of mosquitoes. Similarly, the lack of sanitation can contribute to the formation of small pools of stagnant water, serving as ideal breeding grounds for Aedes. While more prevalent in the tropical and subtropical regions, this viral infection, transmitted by Aedes aegypti and Ae. albopictus mosquitoes, poses a serious public health problem, with outbreaks increasing in number and severity worldwide. In Europe, Aedes mosquito vector species are established in about 22 countries, and dengue fever has been reported for over a decade. In addition to the factors mentioned earlier, the increase in mosquito breeding sites has been identified as a determining factor for the rise in the number of cases.
Consultation on a draft National Gene Drive Policy Guide
28786Commonwealth of Australia (Department of Health and Aged Care), National Gene Technology Scheme, 2024-01-02 13:43:01.
The term gene drive is used to describe organisms which have been genetically modified to increase the rate for a particular trait to spread through a sexually reproducing population, spreading the genes or traits through a species at a faster rate than normal inheritance. An example might be a trait to increase likelihood of offspring to be female and thereby suppress the population of the targeted pest species. The concept of a gene drive is not a new one; these dominant genes – sometimes called selfish genes – are abundant in nature. However, GM gene drive organisms have the potential to be useful for addressing some of the environmental, agricultural, and public health challenges currently faced by Australia, such as conserving native populations, controlling significant exotic pests, or providing public health benefits. As an evolving technology, gene drives may pose risks that are not yet fully understood, including the potential to alter the ecosystem in unpredictable ways. These risks should be acknowledged and managed in a structured and systematic way if Australia wishes to be a future beneficiary of this technology. The Third Review (the Review) of the National Gene Technology Scheme (Scheme), endorsed by all Australian governments on 11 October 2018, recommended “clarifying, and where necessary strengthening, the mechanisms for regulating the environmental release of GM gene drive organisms in Australia” (Recommendation 7b). Review Recommendation 7b and development of the National Gene Drive Policy Guide (Policy Guide) cannot be considered in isolation.
Editorial overview: Conflicts, conflicts everywhere
28768Harmit S Malik, Judith E Mank, Current Opinion in Genetics and Development, 83. 2023-12-31 20:20:14.
Genetic conflicts are pervasive in biology. They arise when genes, sets of genes, or chromosomes enhance their success at significant cost to their genetic neighbors. In some cases, conflicts occur when a ‘cheater’ subverts the rules of genetic inheritance, pitting the interests of the ‘cheater’ against the rest of the genome. In other cases, genetic elements play by the rules of genetic inheritance, but benefit some members of a species at the same time that they harm others. In this issue, we invited a group of scientists at the forefront of the study of different forms of genetic conflicts to share their expertise on some especially intriguing genetic battlegrounds, reflect on their historical underpinnings, summarize current advances, and discuss ideas that will propel the field forward in the years to come. Each of these tales of genetic conflicts is also a tale of evolutionary innovation. Some cases of conflict result from the unique opportunities created by nonfoolproof inheritance rules (e.g. the battle for transmission through gametes or the uniparental inheritance of mitochondria). In other cases, the ‘cheating’ elements (e.g. germline-restricted chromosomes (GRCs)) maintain themselves by creating opportunities to bypass rules their genetic neighbors are forced to abide by. Some conflicts thrive by imposing minimal costs on their neighbors, or even forcing a resolution of conflict so that the interests of all parties are accommodated within the genome. In contrast, others indulge in near-complete chromosomal or gametic fratricide, an observation that is difficult to reconcile without invoking genetic conflicts.
Gene drives, mosquitoes, and ecosystems: An interdisciplinary approach to emerging ethical concerns
28778Ricardo D. Moreno, Luca Valera, Cristián Borgoño, Juan Carlos Castilla, José Luis Riveros, Frontiers in Environmental Science, 11. 2023-12-28 22:14:12.
Gene drives are genetic elements that in sexually reproducing organisms spread faster than those transmitted through a Mendelian fashion. Since gene drives can be engineered to modify different aspects of physiology and reproduction, they have been proposed as a new and revolutionary tool to control vector-borne diseases, particularly those transmitted by the genera Anopheles and Aedes (Culicidae), such as malaria, Dengue and Zika virus. This approach may impact on human health by lowering the transmission of such devastating diseases. However, the release of genetically modified mosquitos (or other species) into the environment raises a series of questions related to the still incipient technology and our present understanding of the complex structure and dynamics of terrestrial and aquatic ecosystems. Moreover, there are ethical concerns about human interventions in natural ecosystems that may eventually impact our way of living or the ecosystems themselves. This work is an interdisciplinary approach that analyzes from a biological, philosophical, and theological perspective the potential ecological impacts on natural environments of the release of genetically modified species, focusing on gene drive-modified mosquitos. It includes theological approach from a Catholic point of view (although it could be easily shared by other Christians) because we hold that world religions give valuable insights even though not everyone may share their groundings. We conclude that the focal problem is the relationship between humans and nature, and the release of genetically modified species may change this relationship unpredictably. However, given the complex interactions in ecosystems, new approaches such as Earth Stewardship principles could provide new and more widely accepted answers involving biological, philosophical, and theological concepts that will help engaging all relevant actors to make a better world.
Antiviral Wolbachia strains associate with Aedes aegypti endoplasmic reticulum membranes and induce lipid droplet formation to restrict dengue virus replication
28757Robson K. Loterio, Ebony A. Monson, Rachel Templin, Jyotika T. de Bruyne, Heather A. Flores, Jason M. Mackenzie, Georg Ramm, Karla J. Helbig, Cameron P. Simmons, Johanna E. Fraser, Applied and Environmental Microbiology, 2023-12-22 16:16:14.
Wolbachia are a genus of insect endosymbiotic bacteria which includes strains wMel and wAlbB that are being utilized as a biocontrol tool to reduce the incidence of Aedes aegypti-transmitted viral diseases like dengue. However, the precise mechanisms underpinning the antiviral activity of these Wolbachia strains are not well defined. Here, we generated a panel of Ae. aegypti-derived cell lines infected with antiviral strains wMel and wAlbB or the non-antiviral Wolbachia strain wPip to understand host cell morphological changes specifically induced by antiviral strains. Antiviral strains were frequently found to be entirely wrapped by the host endoplasmic reticulum (ER) membrane, while wPip bacteria clustered separately in the host cell cytoplasm. ER-derived lipid droplets (LDs) increased in volume in wMel- and wAlbB-infected cell lines and mosquito tissues compared to cells infected with wPip or Wolbachia-free controls. Inhibition of fatty acid synthase (required for triacylglycerol biosynthesis) reduced LD formation and significantly restored ER-associated dengue virus replication in cells occupied by wMel. Together, this suggests that antiviral Wolbachia strains may specifically alter the lipid composition of the ER to preclude the establishment of dengue virus (DENV) replication complexes. Defining Wolbachia’s antiviral mechanisms will support the application and longevity of this effective biocontrol tool that is already being used at scale.
Perspectives of African stakeholders on gene drives for malaria control and elimination: a multi-country survey
28750Finda, M.F., Juma, E.O., Kahamba, N.F. et al., Malaria Journal, 22:8384. 2023-12-21 15:40:51.
Gene drive modified mosquitoes (GDMMs) have the potential to address Africa’s persistent malaria problem, but are still in early stages of development and testing. Continuous engagement of African stakeholders is crucial for successful evaluation and implementation of these technologies. The aim of this multi-country study was, therefore, to explore the insights and recommendations of key stakeholders across Africa on the potential of GDMMs for malaria control and elimination in the continent. A concurrent mixed-methods study design was used, involving a structured survey administered to 180 stakeholders in 25 countries in sub-Saharan Africa, followed by 18 in-depth discussions with selected groups and individuals. Stakeholders were drawn from academia, research and regulatory institutions, government ministries of health and environment, media and advocacy groups. Thematic content analysis was used to identify key topics from the in-depth discussions, and descriptive analysis was done to summarize information from the survey data. Despite high levels of awareness of GDMMs among the stakeholders (76.7%), there was a relatively low-level of understanding of their key attributes and potential for malaria control (28.3%). When more information about GDMMs was provided to the stakeholders, they readily discussed their insights and concerns, and offered several recommendations to ensure successful research and implementation of the technology. These included: (i) increasing relevant technical expertise within Africa, (ii) generating local evidence on safety, applicability, and effectiveness of GDMMs, and (iii) developing country-specific regulations for safe and effective governance of GDMMs. A majority of the respondents (92.9%) stated that they would support field trials or implementation of GDMMs in their respective countries. This study also identified significant misconceptions regarding the phase of GDMM testing in Africa, as several participants incorrectly asserted that GDMMs were already present in Africa, either within laboratories or released into the field. Incorporating views and recommendations of African stakeholders in the ongoing research and development of GDMMs is crucial for instilling stakeholder confidence on their potential application. These findings will enable improved planning for GDMMs in Africa as well as improved target product profiles for the technologies to maximize their potential for solving Africa’s enduring malaria challenge.
Wolbachia infection negatively impacts Drosophila simulans heat tolerance in a strain- and trait-specific manner
28736Liam F Ferguson, Perran A Ross, Belinda van Heerwaarden, bioRxiv, 2023-12-19 16:09:01.
The susceptibility of insects to rising temperatures has largely been measured by their ability to survive thermal extremes. However, until recently, the capacity for maternally inherited endosymbionts to influence insect heat tolerance has been overlooked. Further, the impact of heat on traits like fertility, which can decline at temperatures below the lethal thermal limit has largely been ignored. Here, we assess the impact of three Wolbachia strains (wRi, wAu, and wNo) on the survival and fertility of Drosophila simulans exposed to heat stress during development or as adults. The impact of Wolbachia infection on heat tolerance was generally small and trait/strain specific. Only the wNo infection significantly reduced survival and fertility of adult males after a heat shock. When exposed to a fluctuating heat stress during development, the wRi and wAu strains reduced egg-to-adult survival but only the wNo infection reduced male fertility. Wolbachia densities of all three strains decreased under developmental heat stress, but reductions occurred at temperatures above those that reduced fertility of the host. These findings reveal the complexity of endosymbiont-host-environment interactions and emphasise the necessity to account for endosymbionts and their effect on both survival and fertility when investigating the vulnerability of insects to climate change.
Rapid turnover of pathogen-blocking Wolbachia and their incompatibility loci
28733Shropshire JD, Conner WR, Vanderpool D, Hoffmann AA, Turelli M, Cooper BS., bioRxiv, 2023-12-19 16:02:29.
At least half of all insect species carry maternally inherited Wolbachia alphaproteobacteria, making Wolbachia the most common endosymbionts in nature. Wolbachia spread to high frequencies is often due to cytoplasmic incompatibility (CI), a Wolbachia -induced sperm modification that kills embryos without Wolbachia . Several CI-causing Wolbachia variants, including w Mel from Drosophila melanogaster , also block viruses. Establishing pathogen-blocking w Mel in natural Aedes aegypti mosquito populations has reduced dengue disease incidence, with one study reporting about 85% reduction when w Mel frequency is high. However, w Mel transinfection establishment is challenging in many environments, highlighting the importance of identifying CI-causing Wolbachia variants that stably persist in diverse hosts and habitats. We demonstrate that w Mel-like variants have naturally established in widely distributed holometabolous dipteran and hymenopteran insects that diverged approximately 350 million years ago, with w Mel variants spreading rapidly among these hosts over only the last 100,000 years. Wolbachia genomes contain prophages that encode CI-causing operons ( cifs ). These cifs move among Wolbachia genomes - with and without prophages - even more rapidly than Wolbachia move among insect hosts. Our results shed light on how rapid host switching and horizontal gene transfer contribute to Wolbachia and cif diversity in nature. The diverse w Mel variants we report here from hosts present in different climates offer many new options for broadening Wolbachia -based biocontrol of diseases and pests.
CRISPR/Cas9: a cutting-edge solution for combatting the fall armyworm, Spodoptera frugiperda
28724Gouda, M.N.R., Jeevan, H., Shashank, H.G., Molecular Biology Reports, 51. 2023-12-19 12:15:58.
The utilization of CRISPR/Cas9 in Spodoptera frugiperda, commonly known as fall armyworm, presents a groundbreaking avenue for pest management. With its ability to precisely modify the insect’s genome, CRISPR/Cas9 offers innovative strategies to combat this destructive pest. The application of CRISPR/Cas9 in S. frugiperda holds immense potential. It enables the identification and functional analysis of key genes associated with its behavior, development, and insecticide resistance. This knowledge can unveil novel target sites for more effective and specific insecticides. Additionally, CRISPR/Cas9 can facilitate the development of population control methods by disrupting vital genes essential for survival. However, challenges such as off-target effects and the efficient delivery of CRISPR/Cas9 components remain. Addressing these obstacles is vital to ensure accurate and reliable results. Furthermore, ethical considerations, biosafety protocols, and regulatory frameworks must be integral to the adoption of this technology. Looking forward, CRISPR/Cas9-based gene drive systems hold the potential to promulgate desirable genetic traits within S. frugiperda populations, offering a sustainable and eco-friendly approach. This could curtail their reproductive capabilities or make them more susceptible to certain interventions. In conclusion, CRISPR/Cas9 presents a transformative platform for precise and targeted pest management in S. frugiperda. By deciphering the insect’s genetic makeup and developing innovative strategies, we can mitigate the devastating impact of fall armyworm on agriculture while ensuring environmental sustainability.
Upper Bound on the Mutational Burden Imposed by a CRISPR-Cas9 Gene-Drive Element
28722Michael S. Overton, Sean E. Guy, Xingsen Chen, Alena Martsul, Krypton Carolino, Omar S. Akbari, Justin R. Meyer, Sergey Kryazhimskiy, bioRxiv, 2023-12-19 12:04:20.
CRISPR-Cas9 gene drives (CCGDs) are powerful tools for genetic control of wild populations, useful for eradication of disease vectors, conservation of endangered species and other applications. However, Cas9 alone and in a complex with gRNA can cause double-stranded DNA breaks at off-target sites, which could increase the mutational load and lead to loss of heterozygosity (LOH). These undesired effects raise potential concerns about the long-term evolutionary safety of CCGDs, but the magnitude of these effects is unknown. To estimate how the presence of a CCGD or a Cas9 alone in the genome affects the rates of LOH events and de novo mutations, we carried out a mutation accumulation experiment in yeast Saccharomyces cerevisiae. Despite its substantial statistical power, our experiment revealed no detectable effect of CCGD or Cas9 alone on the genome-wide rates of mutations or LOH events, suggesting that these rates are affected by less than 30%. Nevertheless, we found that Cas9 caused a slight but significant shift towards more interstitial and fewer terminal LOH events, and the CCGD caused a significant difference in the distribution of LOH events on Chromosome V. Taken together, our results show that these genetic elements impose a weak and likely localized additional mutational burden in the yeast model. Although the mutagenic effects of CCGDs need to be further evaluated in other systems, our results suggest that the effect of CCGDs on off-target mutation rates and genetic diversity may be acceptable.
Assessing the sterility and quality of gamma-irradiated pepper weevils, Anthonomus eugenii (Coleoptera: Curculionidae), toward the development of the sterile insect technique
28713Basso, J.V., Labbe, R. and Scott-Dupree, C., Pest Management Science, 2023-12-18 09:48:19.
The pepper weevil (PW), Anthonomus eugenii, is an economically significant pest of cultivated Capsicum spp. pepper crops in North America where it remains a challenge to manage because of its cryptic immature life stages. The sterile insect technique (SIT) is a genetic pest management tactic that relies on the release of insects that have been sterilized with ionizing radiation to lower the population reproductive rate. Toward developing an effective PW-SIT program, this study has, for the first time, investigated the effects of gamma irradiation on the sterility and survival of this species. Among the array of doses tested, we found that pupal PW males and females irradiated at 110 Gy produced no adult offspring. Furthermore, females mated with a male irradiated at 110 Gy had high egg sterility (97.3%), and irradiated females nearly completely failed to lay eggs (97.5%). Individuals irradiated at this dose had a shortened lifespan (lethal time to 50% mortality values of 12 and 11 days for males and females, respectively) and quantitatively reduced spontaneous flight activity. The eclosion rate of PW pupae was not significantly reduced by any radiation treatment. This study suggests that PWs irradiated at a gamma radiation dose of 110 Gy as pupae could feasibly be used in a PW-SIT program, because both males and females were 100% sterile at this dose. These findings will inform the development of a SIT program that could considerably improve the sustainability and effectiveness of PW management in greenhouse and field pepper crops worldwide.
Procesos celulares modificados en Aedes aegypti infectados con Wolbachia y la susceptibilidad al virus dengue
28696López-Ordóñez T, Díaz-Rodarte KI, Torres-Monzón JA, Casas-Martínez M, Danis-Lozano R, Mosso-González C., Salud Pública de México, 2023-12-12 14:35:34.
Objetivo: Analizar la expresión diferencial de proteínas de Aedes aegypti infectados con Wolbachia y su asociación con el ciclo viral del virus dengue (DENV). Material y métodos. Se revisó una base de datos de proteínas de Ae. aegypti infec-tados y no infectados con Wolbachia, cepa wMel y se buscaron estas en revistas indizadas, que hablaran de la proteína y el ciclo viral de DENV. Resultados: La expresión diferencial de proteínas de los mosquitos durante la infección con Wolbachia intervienen en los procesos de entrada, replicación y salida del DENV. Conclusiones: Existen cambios en la expresión de proteínas de células infectadas con Wolbachia, que son necesarias para el ciclo de replicación de DENV, explicando porque algunos mosquitos infectados con Wolbachia son refractarios a la infección por DENV.
Side effects of X-ray irradiation on flight ability of Cydia pomonella moth
28694Huang, S.-W., Zhang, J.-H., Wei, Z.-H., Yang, X.-M., Wang, X.-Y. and Yang, X.-Q., Pest Management Science, 2023-12-12 14:27:29.
The sterile insect technique (SIT) has proven to be an effective approach in managing the population of major invasive pests Our previous studies showed that irradiation of males at a dosage of 366 Gy X-rays resulted in complete sterility. However, the mating competitiveness of sterilized males is significantly compromised, which can be attributed to a decline in their ability to fly. In this study, we examined the flight patterns of both male and female adults of C. pomonella. The results revealed significant variations in the average flight speed of both genders at different stages of maturity, with females displaying longer flight duration and covering greater distances. Effect of irradiation on the flight performance of 3-day-old male moths was further evaluated, as they demonstrated the longest flight distance. The findings indicated a significant decrease in flight distance, duration, and average speed, due to wing deformities caused by irradiation, which also limited the dispersal distance of moths in orchards, as indicated by the mark-and-recapture assay. Quantitative reverse-transcription PCR analysis revealed a down-regulation of flight-related genes such as flightin, myosin heavy chain, and distal-less following radiation exposure. These findings demonstrate that X-ray irradiation at a radiation dose of 366 Gy has a detrimental effect on the flight ability of male C. pomonella adults. These insights not only contribute to a better understanding of how radiation sterilization diminishes the mating competitiveness of male moths, but also aid in the development and improvement of SIT practices for the effectively controlling C. pomonella.
Aerial release of Aedes aegypti male mosquitoes using an unmanned aerial vehicle: a novel control strategy
28690Valdez-Delgado KM, Ríos-Delgado JC, Nettel-Cruz JA, Angulo-Kladt R, Villarreal-Treviño C., Salud Pública de México, 65. 2023-12-12 14:17:22.
Objective. To development of a methodology for the chilling, handling, transport, and release of male Aedes aegypti mosquitoes, reared in insectary conditions to release in the field with unmanned vehicles to compete sexually with wild males in the field. Materials and methods. A population of Ae. aegypti from different areas in Tapachula, Chiapas, was used. Laboratory tests were conducted: Effect of temperature and cooling time on the knockdown, recovery of males, and copulatory success. Results. The chilling temperature of 3 ± 1ºC for 30 min, was used as a knockdown temperature before handling, packing, transportation, and aerial release. The males subjected to the entire process, including the semi-field aerial release test, showed normal sexual behavior activity, obtaining 100% of females inseminated. Conclusion. These results present the feasibility of applying a new control methodology using unmanned aerial vehicle (UAV) as support for the sterile insect release technique (SIT), use of Wolbachia or both, in male Ae. aegypti, for the design of strategies to control their populations.
The Sterile Insect Technique can efficiently reduce the reproduction of the spotted wing drosophila (Drosophila suzukii) in strawberry
28688B. Gard, A. Panel, A. Labbetoul, N. Bosshard, A. Xuereb, B. Cariou, A. Debelle, C. Oliva, S. Fellous, bioRxiv, 2023-12-11 13:19:11.
The spotted wing drosophila (SWD) Drosophila suzukii (Diptera: Drosophilidae) is a highly problematic pest in soft fruit crops. Since its introduction in Europe in 2008, no satisfying nor environment-friendly control method has emerged against this species. The sterile insect technique (SIT) has proven efficient at controlling numerous fruit fly species. In recent years, key elements of SIT against D. suzukii have become available, leading to the publication of encouraging results. However, field- and field-like experiments are notoriously under the influence of various, often unidentified factors granting the need for replicated studies. In this experiment, we assayed the efficacy of a high-performance strain at reducing the reproduction of D. suzukii in complex, yet replicated and controlled conditions. Two ratios of sterile to fertile insects (5:1 and 1:1) using bisexual releases were compared to a control treatment with fertile, wild flies only. The presence of sterile individuals at a 5:1 ratio significantly reduced fly reproduction, measured after 5 days, by an approximate threefold factor. However, the proportion of infested fruits in the treated plots remained unaffected. The number of available berries in the cage appeared as an unexpected determinant of fly infestation, suggesting undocumented density-dependent processes. The success of this assay opens the door to larger scales experiments, over several generations, and, in the near future, the field-evaluation of the efficacy of the SIT to control D. suzukii.
Population suppression with dominant female-lethal alleles is boosted by homing gene drive
28685Jinyu Zhu, Jingheng Chen, Yiran Liu, Xuejiao Xu, Jackson Champer, bioRxiv, 2023-12-07 10:24:25.
Methods to suppress pest insect populations using genetic constructs and repeated releases of male homozygotes have recently been shown to be an attractive alternative to older sterile insect technique based on radiation. Female-specific lethal alleles have substantially increased power, but still require large, sustained transgenic insect releases. Gene drive alleles bias their own inheritance to spread throughout populations, potentially allowing population suppression with a single, small-size release. However, suppression drives often suffer from efficiency issues, and the most well-studied type, homing drives, tend to spread without limit. In this study, we show that coupling female-specific lethal alleles with homing gene drive allowed substantial improvement in efficiency while still retaining the self-limiting nature (and thus confinement) of a lethal allele strategy. Using a mosquito model, we show the required releases sizes for population elimination in a variety of scenarios, including different density growth curves, with comparisons to other systems. Resistance alleles reduced the power of this method, but these could be overcome by targeting an essential gene with the drive while also providing rescue. A proof-of-principle demonstration of this system in Drosophila melanogaster was effective in both basing its inheritance and achieving high lethality among females that inherit the construct in the absence of antibiotic. Overall, our study shows that substantial improvements can be achieved in female-specific lethal systems for population suppression by combining them with a gene drive.
The effect of migration on transmission of Wolbachia in Nilaparvata lugens
28679Zhigang Liu, Tiejun Zhou, Mathematical Biosciences and Engineering, 20. 2023-12-06 13:40:37.
Brown planthopper Nilaparvata lugens, which can transmit rice ragged stunt virus, is a serious and damaging pest to rice plants. Rice plants can protect themselves from the associated diseases of N.lugens by either suppressing or replacing N.lugens by releasing N.lugens infected by a special strain of Wolbachia wStri. The long-distance migration habit of N.lugens is one of the important precursors leading up to the large-scale occurrence of N.lugens. To study the effect of migration on the transmission of Wolbachia in N.lugens, a Wolbachia spreading dynamics model with migration of N.lugens between two patches is put forward. The existence and local stability conditions of equilibrium points of the system and its subsystems are obtained. Moreover, the effects of migration on the dynamic properties and the control of N.lugens are analyzed; the results show that the system can exhibit a bistable phenomenon, and the migration can change the stability of equilibrium infected with wStri from stable to unstable. The quantitative control methods for the migration of the insect N.lugens are proposed, which provide a theoretical guidance for future field experiments. Lastly, we use the Markov chain Monte Carlo (MCMC) method to estimate the parameters of the wild N.lugens migration model based on limited observational data; the numerical simulation results show that migration can increase the quantity of N.lugens, which is consistent with the relevant experimental results.
Transformative Approaches for Sustainable Weed Management: The Power of Gene Drive and CRISPR-Cas9
28677Kumam, Y.; Trick, H.N.; Vara Prasad, P.V.; Jugulam, M., Genes, 14. 2023-12-06 13:31:23.
Weeds can negatively impact crop yields and the ecosystem’s health. While many weed management strategies have been developed and deployed, there is a greater need for the development of sustainable methods for employing integrated weed management. Gene drive systems can be used as one of the approaches to suppress the aggressive growth and reproductive behavior of weeds, although their efficacy is yet to be tested. Their popularity in insect pest management has increased, however, with the advent of CRISPR-Cas9 technology, which provides specificity and precision in editing the target gene. This review focuses on the different types of gene drive systems, including the use of CRISPR-Cas9-based systems and their success stories in pest management, while also exploring their possible applications in weed species. Factors that govern the success of a gene drive system in weeds, including the mode of reproduction, the availability of weed genome databases, and well-established transformation protocols are also discussed. Importantly, the risks associated with the release of weed populations with gene drive-bearing alleles into wild populations are also examined, along with the importance of addressing ecological consequences and ethical concerns.
Viruses that ‘infect’ viruses: Cas12f1 and Cas9 gene drive in HSV1
28673Hongsheng Dai, Qiaorui Yao, Zhuangjie Lin, Keyuan Lai, Xianyin Zeng, Guangxiong Lei, Tongwen Zhang, bioRxiv, 2023-12-06 12:08:32.
Cas9-based synthetic gene drives constitute some minimal elements capable of editing DNA with sequence specificity. However, they face high resistance rate and mitigation strategies developed so far are difficult to implement. Here, we engineered herpes simplex virus type 1 (HSV1) as a vector for gene drive, generated viruses carrying either Cas9 or Cas12f1 gene drives, and compared their performance in spreading the gene drive and causing resistance. Based on phenotypical staining and NGS sequencing, we showed that HSV1 supported fast and effective transmission of gene drive, and Cas12f1 gene drive yielded higher conversion and lower resistance than Cas9 gene drive. Cas12f1 gene drive is a promising alternative to Cas9 gene drive, and HSV1 is a reliable and fast platform to evaluate gene drive. Gene drive viruses worked like pathogens that specifically infect viruses and could potentially be used to attenuate viral infection.
A population modification gene drive targeting both Saglin and Lipophorin impairs Plasmodium transmission in Anopheles mosquitoes
28671Emily I Green, Etienne Jaouen, Dennis Klug, Roenick Proveti Olmo, Amandine Gautier, Stéphanie Blandin, Eric Marois, eLife, 12. 2023-12-06 11:55:36.
Lipophorin is an essential, highly expressed lipid transport protein that is secreted and circulates in insect hemolymph. We hijacked the Anopheles coluzzii Lipophorin gene to make it co-express a single-chain version of antibody 2A10, which binds sporozoites of the malaria parasite Plasmodium falciparum. The resulting transgenic mosquitoes show a markedly decreased ability to transmit Plasmodium berghei expressing the P. falciparum circumsporozoite protein to mice. To force the spread of this anti-malarial transgene in a mosquito population, we designed and tested several CRISPR/Cas9-based gene drives. One of these is installed in, and disrupts, the pro-parasitic gene Saglin and also cleaves wild-type Lipophorin, causing the anti-malarial modified Lipophorin version to replace the wild type and hitch-hike together with the Saglin drive. Although generating drive-resistant alleles and showing instability in its gRNA-encoding multiplex array, the Saglin-based gene drive reached high levels in caged mosquito populations and efficiently promoted the simultaneous spread of the antimalarial Lipophorin::Sc2A10 allele. This combination is expected to decrease parasite transmission via two different mechanisms. This work contributes to the design of novel strategies to spread antimalarial transgenes in mosquitoes, and illustrates some expected and unexpected outcomes encountered when establishing a population modification gene drive.
Homing-based gene drives can introgress rapidly into local genetic backgrounds with minimal chromosomal conversion in Anopheles gambiae
28663Tony Nolan, Poppy Pescod, Giulia Bevivino et al., Research Square, 2023-12-06 10:02:37.
CRISPR-Cas9 gene drive control strategies use a homing selfish genetic element which induces a double-stranded break at the target site and is copied into the opposing chromosome, breaking an essential gene and ensuring super-Mendelian inheritance. During the copying process SNPs could potentially be transferred from one chromosome to another, converting tracts of the target to match the recipient. This would reduce the ability of gene drives to escape their genetic background of origin, with potential impacts on carrier fitness and the success of the strategy. In this study we use Anopheles gambiae strains with variation around the target site to determine the extent of chromosomal conversion during gene drive activity and the mechanism of inheritance bias. We confirm a homing-based mechanism and show that two thirds of homing events are resolved within 50 bp, showing that gene drive introgression into a wild strain can occur within one generation.
Germline transformation of the West Nile Virus and avian malaria vector Culex quinquefasciatus Say using the piggyBac transposon system
28653Katherine Nevard, Rajdeep Kaur, Tim Harvey-Samuel, bioRxiv, 2023-12-05 11:04:04.
Culex quinquefasciatus Say is a mosquito which acts as a vector for numerous diseases including West Nile Virus, lymphatic filariasis and avian malaria, over a broad geographical range. As the effectiveness of insecticidal mosquito control methods declines, the need has grown to develop genetic control methods to curb the spread of disease. The piggyBac transposon system - the most widely used genetic transformation tool in insects, including mosquitoes - generates quasi-random insertions of donor DNA into the host genome. However, despite the broad reported species range of piggyBac, previous attempts to use this tool to transform Culex quinquefasciatus mosquitoes have failed. Here we report the first successful transformation of Culex quinquefasciatus with the piggyBac transposon system. Using commercially synthesised piggyBac mRNA as a transposase source, we were able to generate three independent insertions of a ZsGreen fluorescent marker gene, with transformation efficiencies of up to 5%. Through this work, we have expanded the genetic toolkit available for the genetic manipulation of Culex mosquitoes and thus removed a barrier to developing novel genetic control methods in this important disease vector.
A homing rescue gene drive with multiplexed gRNAs reaches high frequency in cage populations but generates functional resistance
28651Jingheng Chen, Shibo Hou, Ruobing Feng, Xuejiao Xu, Nan Liang, Jackson Champer, bioRxiv, 2023-12-05 10:59:48.
CRISPR homing gene drive is a potent technology with considerable potential for managing populations of medically and agriculturally significant insects. It induces a bias in the inheritance of the drive allele in progeny, rapidly spreading desired genes throughout the population. Homing drives operate by Cas9 cleavage followed by homology-directed repair, copying the drive allele to the wild-type chromosome. However, resistance alleles formed by end-joining repair pose a significant obstacle to the spread of the drive. To address this challenge, we created a homing drive targeting the essential but haplosufficient hairy gene. Our strategy involves spreading the drive construct through the homing process, eliminating nonfunctional resistance, which are recessive lethal, while rescuing drive-carrying individuals with a recoded version of hairy. This strategy eliminates resistance more slowly than a previous strategy targeting haplolethal genes, but it may be easier to construct in non-model organisms. Our drive inheritance rate was moderate, and multigenerational cage studies showed quick drive spread to 96-97% of the population. However, the drive failed to reach the whole population due to the formation of functional resistance alleles, despite use of four gRNAs, a strategy that previously was successful at preventing functional resistance. Sequencing showed that these alleles had a large deletion and must have utilized an alternate start codon. The resistance allele had a modest fitness advantage over the drive in a cage study, which could prevent long-term persistence of the drive, especially if cargo genes had an additional fitness cost. Thus, revised design strategies targeting more essential regions of a target gene may often be necessary to avoid such functional resistance, even when using multiplexed gRNAs.
Reduced dengue incidence following city-wide wMel Wolbachia mosquito releases throughout three Colombian cities: Interrupted time series analysis and a prospective case-control study
28647Velez ID, Tanamas SK, Arbelaez MP, Kutcher SC, Duque SL, Uribe A, et al., PLoS Neglected Tropical Diseases, 2023-12-05 10:26:43.
The introduction of Wolbachia (wMel strain) into Aedes aegypti mosquitoes reduces their capacity to transmit dengue and other arboviruses. Randomised and non-randomised studies in multiple countries have shown significant reductions in dengue incidence following field releases of wMel-infected Ae. aegypti. Stable introduction of wMel into local Ae. aegypti populations was associated with a significant and sustained reduction in dengue incidence across three Colombian cities. These results from the largest contiguous Wolbachia releases to-date demonstrate the real-world effectiveness of the method across large urban populations and, alongside previously published results, support the reproducibility of this effectiveness across different ecological settings.
Large-scale releases and establishment of wMel Wolbachia in Aedes aegypti mosquitoes throughout the Cities of Bello, Medellín and Itagüí, Colombia
28645Velez ID, Uribe A, Barajas J, Uribe S, Ángel S, Suaza-Vasco JD, et al., PLoS Neglected Tropical Diseases, 2023-12-05 10:11:27.
The wMel strain of Wolbachia has been successfully introduced into Aedes aegypti mosquitoes and has been shown to reduce the transmission of dengue and other Aedes-borne viruses. Here we report the entomological results from phased, large-scale releases of Wolbachia infected Ae. aegypti mosquitoes throughout three contiguous cities located in the Aburrá Valley, Colombia. These results, from the largest contiguous releases of wMel Wolbachia mosquitoes to date, highlight the operational feasibility of implementing the method in large urban settings. Based on results from previous studies, we expect that Wolbachia establishment will be sustained long term. Ongoing monitoring will confirm Wolbachia persistence in local mosquito populations and track its establishment in the remaining areas.
Modelling and analysis of periodic impulsive releases of the Nilaparvata lugens infected with wStri-Wolbachia
28638Xiangjun Dai, Qi Quan and Jianjun Jiao, Journal of Biological Dynamics, 17:1. 2023-12-04 12:23:12.
In this paper, we formulate a population suppression model and a population replacement model with periodic impulsive releases of Nilaparvata lugens infected with wStri. The conditions for the stability of wild-N.lugens -eradication periodic solution of two systems are obtained by applying the Floquet theorem and comparison theorem. And the sufficient conditions for the persistence in the mean of wild N.lugens are also given. In addition, the sufficient conditions for the extinction and persistence of the wild N.lugens in the subsystem without wLug are also obtained. Finally, we give numerical analysis which shows that increasing the release amount or decreasing the release period are beneficial for controlling the wild N.lugens, and the efficiency of population replacement strategy in controlling wild populations is higher than that of population suppression strategy under the same release conditions.
Food for thought: Assessing the consumer welfare impacts of deploying irreversible, landscape-scale biotechnologies
28628Michael S. Jones, Zachary S. Brown, Food Policy, 121. 2023-12-04 10:24:17.
Genetically engineered insects have gained attention as regionally deployed pest control technologies, with substantial applications in agriculture for combatting intractable crop pests and diseases. One potential tool is a ‘gene drive’, using CRISPR-based gene editing. In gene drive, preferentially inherited, engineered traits are spread throughout a geographic area to reduce pest populations or inhibit disease transmission, while also potentially reducing pesticide use and crop prices. But the self-perpetuating nature of gene drives presents a consequence, in that consumers could eventually be limited to only host crops grown in the presence of these genetically engineered insects. In this study, we analyze potential consumer welfare impacts of these technologies using discrete choice experiment data from a representative sample of U.S. adults, examining preferences regarding gene drive use to control spotted wing drosophila in blueberries and Asian citrus psyllid in orange juice (OJ) production. We find smaller average discounts for gene drives versus increased conventional pesticide use or genetically modified crops. Only 27% and 25% of blueberry and OJ consumers, respectively, are estimated to derive disutility from gene drives. However, gene drive disutility for these consumers is so large that elimination of non-drive options from their choice sets results in negative (blueberries) or neutral (OJ) effects to aggregate consumer welfare when weighed against gains to other consumers from reduced prices. Positive welfare effects are recovered by retaining availability of non-gene-drive products. We argue that this type of analysis will be increasingly important as landscape-level biotechnologies are deployed to address challenges to agricultural sustainability.
Manipulating the Destiny of Wild Populations Using CRISPR
28626Raban R, Marshall JM, Hay BA, Akbari OS., Annual Reviews, 57:361-390. 2023-12-04 09:57:57.
Genetic biocontrol aims to suppress or modify populations of species to protect public health, agriculture, and biodiversity. Advancements in genome engineering technologies have fueled a surge in research in this field, with one gene editing technology, CRISPR, leading the charge. This review focuses on the current state of CRISPR technologies for genetic biocontrol of pests and highlights the progress and ongoing challenges of using these approaches.
Anopheles gambiae on remote islands in the Indian Ocean: origins and prospects for malaria elimination by genetic modification of extant populations
28597Ditter, R.E., Campos, M., Crepeau, M.W. et al., Scientific Reports, 13. 2023-11-29 16:40:50.
The mosquito Anopheles gambiae s.s. is a primary malaria vector throughout sub-Saharan Africa including the islands of the Comoros archipelago (Anjouan, Grande Comore, Mayotte and Mohéli). These islands are located at the northern end of the Mozambique Channel in eastern Africa. Previous studies have shown a relatively high degree of genetic isolation between the Comoros islands and mainland populations of A. gambiae, but the origin of the island populations remains unclear. Here, we analyzed phylogenetic relationships among island and mainland populations using complete mitochondrial genome sequences of individual A. gambiae specimens. This work augments earlier studies based on analysis of the nuclear genome. We investigated the source population of A. gambiae for each island, estimated the number of introductions, when they occurred and explored evidence for contemporary gene flow between island and mainland populations. These studies are relevant to understanding historical patterns in the dispersal of this important malaria vector and provide information critical to assessing their potential for the exploration of genetic-based vector control methods to eliminate this disease. Phylogenetic analysis and haplotype networks were constructed from mitogenome sequences of 258 A. gambiae from the four islands. In addition, 112 individuals from seven countries across sub-Saharan Africa and Madagascar were included to identify potential source populations. Our results suggest that introduction events of A. gambiae into the Comoros archipelago were rare and recent events and support earlier claims that gene flow between the mainland and these islands is limited. This study is concordant with earlier work suggesting the suitability of these oceanic islands as appropriate sites for conducting field trial releases of genetically engineered mosquitoes (GEMs).
Wolbachia Transinfection Via Embryonic Microinjection
28593Zhang, M., Xi, Z., Methods in Molecular Biology, 2739. 2023-11-29 16:22:25.
The process of transferring Wolbachia from one species to another to establish a stable, maternally inherited infection in the target species is known as transinfection. The success of transinfection is primarily achieved through embryonic microinjection, which is the most direct and efficient means of delivering Wolbachia into the germline of the target species and establishing stable maternal transmission. For the fundamental studies, transinfection is often used to characterize Wolbachia-host interactions, including Wolbachia host range, the role of host or bacterial factors in symbiosis, and evolution of Wolbachia-host associations. For the applied studies, use of transinfection to generate a novel infection in the target species is the first step to build the weapon for both population replacement and population suppression for controlling insect pests or their transmitted diseases. For the primary dengue vector Aedes aegypti and Anopheles vectors of malaria, which either do not naturally carry Wolbachia or are infected with strains that lack necessary features for implementation, transinfection can be established by introducing a novel strain capable of inducing both cytoplasmic incompatibility (CI) and pathogen blocking. For A. albopictus and Culex mosquito species, which naturally harbor CI-inducing Wolbachia, transinfection can be achieved by either introducing a novel strain to generate superinfection or replacing the native infection with a different Wolbachia strain in a symbiont-free line, which is derived from antibiotic treatment of the wild type. Here, we use A. aegypti as an example to describe the Wolbachia transinfection method, which can be adapted to other insect species, such as planthoppers, according to their specific developmental requirements.
Measuring Host Fitness Effects and Transmission of Wolbachia Strains in Aedes aegypti Mosquitoes
28587Ross, P.A., Methods in Molecular Biology, 2739. 2023-11-29 15:49:58.
Lines of Aedes aegypti mosquitoes infected with heritable Wolbachia bacteria are being developed and released for arbovirus control. Coordinated releases of lab-reared Wolbachia mosquitoes have reduced local disease incidence by spreading virus-blocking Wolbachia strains and by crashing mosquito populations through incompatible male releases. The phenotypic effects of Wolbachia are diverse and depend on both genetics and the environment. Accurate assessments of Wolbachia effects in mosquitoes are essential, as such effects can make the difference between success and failure of a Wolbachia release program. This chapter provides guidelines for testing key Wolbachia host effects and transmission in Aedes aegypti: the most important arbovirus vector and the most common target of Wolbachia release programs. The protocols should be useful for evaluating mosquito strains prior to field release.
Procedures for the Detection of Wolbachia-Conferred Antiviral Protection in Drosophila melanogaster
28581Chrostek, E., Methods in Molecular Biology, 2739. 2023-11-29 15:34:15.
Spread of Wolbachia infections in host populations may be enhanced by Wolbachia-conferred protection from viral pathogens. Wolbachia-infected Drosophila melanogaster survive the pathogenic effects of positive-sense single-stranded RNA virus infections at a higher rate than the flies without Wolbachia. The protection can occur with or without detectable reduction in virus titer. For the comparisons to be meaningful, Wolbachia-harboring and Wolbachia-free insects need to be genetically matched, and original populations of gut microbiota need to be restored after the removal of Wolbachia using antibiotics. Here, I describe the procedures needed to detect Wolbachia-conferred antiviral protection against Drosophila C virus measured as the difference in survival and viral titer between flies with and without Wolbachia.
Use of Drosophila Transgenics to Identify Functions for Symbiont Effectors
28574Cortez, C.T., Murphy, R.O., Owens, I.M., Beckmann, J.F., Methods in Molecular Biology, 2739. 2023-11-29 14:35:56.
Wolbachia, one of the most successful and studied insect symbionts, and Drosophila, one of the most understood model insects, can be exploited as complementary tools to unravel mechanisms of insect symbiosis. Although Wolbachia itself cannot be grown axenically as clonal isolates or genetically manipulated by standard methods, its reproductive phenotypes, including cytoplasmic incompatibility (CI), have been elucidated using well-developed molecular tools and precise transgenic manipulations available for Drosophila melanogaster. Current research only scratches the surface of how Drosophila can provide a tool for understanding Wolbachia’s evolutionary success and the molecular roles of its genetic elements. Here, we briefly outline basic methodologies inherent to transgenic Drosophila systems that have already contributed significant advances in understanding CI, but may be unfamiliar to those who lack experience in Drosophila genetics. In the future, these approaches will continue providing significant insights into Wolbachia that undoubtedly will be extended to other insect symbionts and their biological capabilities.
