Scholarly Literature

This is a database of scholarly literature that concentrates currently on natural and engineered selfish genetic elements (gene drives).  The latest are shown here.
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Toward a CRISPR-Cas9-Based Gene Drive in the Diamondback Moth Plutella xylostella

24426
X. Xu, T. Harvey-Samuel, H. A. Siddiqui, J. X. D. Ang, M. E. Anderson, C. M. Reitmayer, E. Lovett, P. T. Leftwich, M. You and L. Alphey,  The CRISPR Journal,  5:224-236. 2022-04-01 10:25:39.
Promising to provide powerful genetic control tools, gene drives have been constructed in multiple dipteran insects, yeast, and mice for the purposes of population elimination or modification. However, it remains unclear whether these techniques can be applied to lepidopterans. Here, we used endogenous regulatory elements to drive Cas9 and single guide RNA (sgRNA) expression in the diamondback moth (DBM), Plutella xylostella, and test the first split gene drive system in a lepidopteran. The DBM is an economically important global agriculture pest of cruciferous crops and has developed severe resistance to various insecticides, making it a prime candidate for such novel control strategy development. A very high level of somatic editing was observed in Cas9/sgRNA transheterozygotes, although no significant homing was revealed in the subsequent generation. Although heritable Cas9-medated germline cleavage as well as maternal and paternal Cas9 deposition were observed, rates were far lower than for somatic cleavage events, indicating robust somatic but limited germline activity of Cas9/sgRNA under the control of selected regulatory elements. Our results provide valuable experience, paving the way for future construction of gene drives or other Cas9-based genetic control strategies in DBM and other lepidopterans.

Overview of paratransgenesis as a strategy to control pathogen transmission by insect vectors

21579
N. A. Ratcliffe, J. P. Furtado Pacheco, P. Dyson, H. C. Castro, M. S. Gonzalez, P. Azambuja and C. B. Mello,  Parasites and Vectors,  15:112. 2022-03-31 12:47:27.
This article presents an overview of paratransgenesis as a strategy to control pathogen transmission by insect vectors. It first briefly summarises some of the disease-causing pathogens vectored by insects and emphasises the need for innovative control methods to counter the threat of resistance by both the vector insect to pesticides and the pathogens to therapeutic drugs. Subsequently, the state of art of paratransgenesis is described, which is a particularly ingenious method currently under development in many important vector insects that could provide an additional powerful tool for use in integrated pest control programmes. The requirements and recent advances of the paratransgenesis technique are detailed and an overview is given of the microorganisms selected for genetic modification, the effector molecules to be expressed and the environmental spread of the transgenic bacteria into wild insect populations. The results of experimental models of paratransgenesis developed with triatomines, mosquitoes, sandflies and tsetse flies are analysed. Finally, the regulatory and safety rules to be satisfied for the successful environmental release of the genetically engineered organisms produced in paratransgenesis are considered.

Comparison of Ground Release and Drone-Mediated Aerial Release of Aedes aegypti Sterile Males in Southern Mexico: Efficacy and Challenges

21574
C. F. Marina, P. Liedo, J. G. Bond, A. R. Osorio, J. Valle, R. Angulo-Kladt, Y. Gómez-Simuta, I. Fernández-Salas, A. Dor and T. Williams,  Insects,  13. 2022-03-31 12:43:15.
Sterile males of Aedes aegypti were released once a week for 8 weeks to evaluate the dispersal efficiency of ground and aerial drone release methods in a rural village of 26 Ha in southern Mexico. Indoor and outdoor BG-Sentinel traps were placed in 13–16 houses distributed throughout the village. The BG traps were activated 48 h after the release of the sterile males and functioned for a 24 h period following each release. Over the 8-week period of simultaneous ground and aerial releases, an average of 85,117 ± 6457 sterile males/week were released at ground level and 86,724 ± 6474 sterile males/week were released using an aerial drone. The ground release method resulted in higher numbers of captured males (mean = 5.1 ± 1.4, range 1.1–15.7 sterile males/trap) compared with the aerial release method (mean = 2.6 ± 0.8, range 0.5–7.3 sterile males/trap) (p < 0.05). Similarly, the prevalence of traps that captured at least one sterile male was significantly higher for ground release compared to the aerial release method (p < 0.01). The lower numbers of sterile males captured in the aerial release method could be due to mortality or physical injury caused by the chilling process for immobilization, or the compaction of these insects during transport and release. However, aerial releases by a two-person team distributed insects over the entire village in just 20 min, compared to ~90 min of work for a five-person team during the ground release method. Ground release also resulted in higher aggregations of males and some villagers reported feeling discomfort from the presence of large numbers of mosquitoes in and around their houses. We conclude that modifications to the handling and transport of sterile males and the design of containers used to store males are required to avoid injury and to improve the efficiency of aerial releases for area-wide SIT-based population suppression programs targeted at mosquito vectors of human disease.

Genetic Stability and Fitness of Aedes aegypti Red-Eye Genetic Sexing Strains With Pakistani Genomic Background for Sterile Insect Technique Applications

22452
M. Misbah-ul-Haq, D. O. Carvalho, L. D. de la Fuente, A. A. Augustinos and K. Bourtzis,  Frontiers in Bioengineering and Biotechnology,  10. 2022-03-31 09:37:56.
The mosquito species Aedes aegypti is the primary transmitter of viruses that cause endemic diseases like dengue in Pakistan. It is also a cause of other vector-borne diseases like yellow fever, Zika fever, and chikungunya, which significantly impact human health worldwide. In the absence of efficient vaccines (except for yellow fever) or drugs, vector control methods, such as the sterile insect technique (SIT), have been proposed as additional tools for the management of these diseases. Mosquito SIT programs are based on the release of sterile males and it is important female releases to be ideally zero or to be kept at a minimum, since females are the ones that bite, blood-feed and transmit pathogens. Recently, an Ae. aegypti genetic sexing strain (GSS), with and without a recombination-suppressing inversion (Inv35), was developed using the eye color as a selectable marker, with males having black eyes and females red eyes. In the present study, we introgressed the sexing features and the Inv35 of the Ae. aegypti red-eye GSS into the Pakistani genomic background aiming to their future use for SIT applications in the country. Both introgressed strains, the Red-eye GSS-PAK and the Red-eye GSS/Inv35-PAK, were evaluated in respect to their genetic stability and biological quality by assessing parameters like recombination rate, fecundity, fertility, pupal and adult recovery, time of development, pupal weight, survival, and flight ability in comparison with a wild Pakistani population (PAK). The results suggest that the sexing features and the recombination suppression properties of Inv35 were not affected after their introgression into the local genomic background; however, some biological traits of the two newly constructed strains were affected, positively or negatively, suggesting that a thorough quality control analysis should be performed after the introgression of a GSS into a new genomic background prior to its use in SIT field trials or applications. The importance of using GSS with local genomic background for SIT applications against Aedes aegypti is also discussed.

Prescribing engagement in environmental risk assessment for gene drive technology

21689
S. Hartley, A. Kokotovich and C. McCalman,  Regulation and Governance,  2022-03-29 08:17:38.
Abstract Gene drive technology is a nascent biotechnology with the potential to purposefully alter or eliminate a species. There have been broad calls for engagement to inform gene drive governance. Over the past seven years, the gene drive community has been developing risk assessment guidelines to determine what form future gene drive risk assessments take, including whether and how they involve engagement. To explore who is developing these guidelines and how engagement in risk assessment is being prescribed, we conduct a document analysis of gene drive risk assessment guideline documents from 2014 to 2020. We found that a narrow set of organizations have developed 10 key guideline documents and that with only one exception the documents prescribe a narrow, vague, or completely absent role for engagement in gene drive risk assessment. Without substantively prescribed engagement in guidelines, the relevance, rigor, and trustworthiness of gene drive risk assessment and governance will suffer.

Mathematical modelling to assess the feasibility of Wolbachia in malaria vector biocontrol

22650
S. Andreychuk and L. Yakob,  Journal of Theoretical Biology,  542. 2022-03-29 06:30:06.
Releasing mosquitoes transinfected with the endosymbiotic bacterium Wolbachia is a novel strategy for interrupting vector-borne pathogen transmission. Following its success in controlling arboviruses spread by Aedes aegypti, this technology is being adapted for anopheline malaria vectors. However, antagonistic interactions between Wolbachia and naturally resident Asaia bacteria in malaria vectors have been demonstrated experimentally, potentially jeopardising Wolbachia biocontrol. We developed the first mathematical model accounting for interspecific competition between endosymbionts to assess the feasibility of this novel strategy for controlling malaria. First, Asaia prevalences among natural mosquito populations were compared with simulations parametrized with rates of Asaia transmission reported from laboratory studies. Discrepancies between projections and natural Asaia prevalences indicated potential overestimation of Asaia transmissibility in artificial laboratory settings. With parametrization that matches natural Asaia prevalence, simulations identified redundancies in Asaia's many infection routes (vertical, sexual and environmental). This resilience was only overcome when Wolbachia conferred very high resistance to environmental infection with Asaia, resulting in Wolbachia fixation and Asaia exclusion. Wolbachia's simulated spread was prevented when its maternal transmission was impeded in coinfected mosquitoes and the pre-control Asaia prevalence was beyond a threshold of 60-75%. This theoretical assessment highlights critical next steps in laboratory experiments to inform this strategy's feasibility. (c) 2022 The Author(s). Published by Elsevier Ltd.

Finding the strongest gene drive: Simulations reveal unexpected performance differences between Anopheles homing suppression drive candidates

21566
S. E. Champer, I. K. Kim, A. G. Clark, P. W. Messer and J. Champer,  bioRxiv,  2022.03.28.486009. 2022-03-28 12:29:28.
Recent experiments have produced several Anopheles gambiae homing gene drives that disrupt female fertility genes, thereby eventually inducing population collapse. Such drives may be highly effective tools to combat malaria. One such homing drive, based on the zpg promoter driving CRISPR/Cas9, was able to eliminate a cage population of mosquitoes. A second version, purportedly improved upon the first by incorporating an X-shredder element (which biases inheritance towards male offspring), was similarly successful. Here, we re-analyze the data of each of these gene drives and suggest an alternative interpretation of their performance. We assess each suppression drive within an individual-based simulation framework that models mosquito population dynamics in continuous space. We find that the combined homing/X-shredder drive is actually less effective at population suppression within the context of our mosquito population model. In particular, the combined drive often fails to completely suppress the population, instead resulting in an unstable equilibrium between drive and wild-type alleles. By contrast, otherwise similar drives based on the nos promoter may prove to be more promising candidates for future development due to potentially superior performance.Competing Interest StatementThe authors have declared no competing interest.

Cytoplasmic incompatibility: A Wolbachia toxin–antidote mechanism comes into view

21546
M. Hochstrasser,  Current Biology,  32:R287-R289. 2022-03-28 11:56:01.
The Wolbachia cidA and cidB genes promote bacterial endosymbiont inheritance through the host female germline. CidB is now shown to load into maturing sperm nuclei. Following fertilization, it disrupts paternal chromosome condensation, triggering embryonic arrest if not countered by CidA in Wolbachia-infected eggs.

B-A Chromosome Translocations Possessing an A Centromere Partly Overcome the Root-Restricted Process of Chromosome Elimination in Aegilops speltoides

21726
D. Li, A. Ruban, J. Fuchs, H. Kang and A. Houben,  Frontiers in Cell and Developmental Biology,  10. 2022-03-28 09:37:44.
Some eukaryotes exhibit dramatic genome size differences between cells of different organs, resulting from the programmed elimination of chromosomes. Aegilops speltoides is an annual diploid species from the Poaceae family, with a maximum number of eight B chromosomes (Bs) in addition to its inherent seven pairs of standard A chromosomes (As). The Bs of this species undergo precise elimination in roots early in embryo development. In areal parts of the plant, the number of Bs is stable. To affect the root restricted process of B chromosome elimination, we employed X-ray mutagenesis, and different types of restructured Bs were identified. Standard Bs were observed in all analyzed shoots of mutagenized plants, while B-A translocations were only observed in 35.7% of F1 plants. In total 40 different B variants inconsistently escaped the elimination process in roots. As a result, mosaicism of B chromosome variants was found in roots. Only a small B chromosome fragment fused to an A chromosome was stably maintained in roots and shoots across F1 to F3 generations. The absence of B-A translocation chromosomes possessing a derived B centromere in root cells implies that the centromere of the B is a key component of the chromosome elimination process.

Field Suppression of Spotted Wing Drosophila (SWD) (Drosophila suzukii Matsumura) Using the Sterile Insect Technique (SIT)

21241
R. A. Homem, Z. Mateos-Fierro, R. Jones, D. Gilbert, A. R. McKemey, G. Slade and M. T. Fountain,  Insects,  13. 2022-03-26 06:14:15.
Drosophila suzukii (spotted wing drosophila—SWD) is an economically important pest of soft and stone fruit worldwide. Control relies on broad-spectrum insecticides, which are neither fully effective nor environmentally sustainable. The sterile insect technique (SIT) is a proven, effective and environmentally friendly pest-management tool. Here, we investigated, for the first time, the potential of using SIT to control D. suzukii in field conditions without physical barriers that limit insect invasion. A proprietary method of rearing and irradiation with X-rays was used to obtain males that were 99% sterile. Sterile males were released twice per week from April to October 2021 on a site in Kent, UK, where everbearing strawberries were grown in open polytunnels. The infestation of wild female D. suzukii was monitored weekly using red sticky traps with dry lure at the treated site and at two similar control sites that did not receive sterile male releases. Releases of sterile males suppressed the wild female D. suzukii population by up to 91% in comparison with the control sites. We thus demonstrated the feasibility of SIT to achieve season-long control of D. suzukii using early, sustained and dynamically targeted releases of sterile males. This provides a promising environmentally friendly method to control this important pest.

Evolution of eukaryotic centromeres by drive and suppression of selfish genetic elements

21559
T. Kumon and M. A. Lampson,  Seminars in Cell and Developmental Biology,  2022-03-25 12:12:40.
Despite the universal requirement for faithful chromosome segregation, eukaryotic centromeres are rapidly evolving. It is hypothesized that rapid centromere evolution represents an evolutionary arms race between selfish genetic elements that drive, or propagate at the expense of organismal fitness, and mechanisms that suppress fitness costs. Selfish centromere DNA achieves preferential inheritance in female meiosis by recruiting more effector proteins that alter spindle microtubule interaction dynamics. Parallel pathways for effector recruitment are adaptively evolved to suppress functional differences between centromeres. Opportunities to drive are not limited to female meiosis, and selfish transposons, plasmids and B chromosomes also benefit by maximizing their inheritance. Rapid evolution of selfish genetic elements can diversify suppressor mechanisms in different species that may cause hybrid incompatibility.

Opening up, closing down, or leaving ajar? How applications are used in engaging with publics about gene drive

21287
A. W. Russell, A. Stelmach, S. Hartley, L. Carter and S. Raman,  Journal of Responsible Innovation,  2022-03-22 12:32:50.
Public engagement and responsible innovation are strongly emphasised in gene drive research, together with the goal of addressing societal challenges, notably, malaria and environmental conservation. We aim to explore whether public engagement is used to ‘open up' or ‘close down' opportunities to shape gene drive research. Drawing on interviews with gene drive developers and stakeholders, we investigate how the public communication of gene drive is conceived. We find that traditional closing-down tendencies remain, but that there are new and encouraging opening-up approaches. Consistent with responsible innovation thinking, these frame gene drive as multifaceted, context-dependent and requiring deeper deliberation. We also identify a third ‘leaving ajar’ approach that seeks to engage with and respond to local communities and modify technological applications to be more acceptable. Innovation system constraints may well temper current aspirations to open up; framing public conversations around understandings of public good could offer a way forward.

CRISPR-mediated knockout of cardinal and cinnabar eye pigmentation genes in the western tarnished plant bug

21243
C. C. Heu, R. J. Gross, K. P. Le, D. M. LeRoy, B. Fan, J. J. Hull, C. S. Brent and J. A. Fabrick,  Scientific Reports,  12. 2022-03-22 06:19:14.
The western tarnished plant bug, Lygus hesperus, is a key hemipteran pest of numerous agricultural, horticultural, and industrial crops in the western United States and Mexico. A lack of genetic tools in L. hesperus hinders progress in functional genomics and in developing innovative pest control methods such as gene drive. Here, using RNA interference (RNAi) against cardinal (LhCd), cinnabar (LhCn), and white (LhW), we showed that knockdown of LhW was lethal to developing embryos, while knockdown of LhCd or LhCn produced bright red eye phenotypes, in contrast to wild-type brown eyes. We further used CRISPR/Cas9 (clustered regularly interspaced palindromic repeats/CRISPR-associated) genome editing to generate germline knockouts of both LhCd (Card) and LhCn (Cinn), producing separate strains of L. hesperus characterized by mutant eye phenotypes. Although the cardinal knockout strain Card exhibited a gradual darkening of the eyes to brown typical of the wild-type line later in nymphal development, we observed bright red eyes throughout all life stages in the cinnabar knockout strain Cinn, making it a viable marker for tracking gene editing in L. hesperus. These results provide evidence that CRISPR/Cas9 gene editing functions in L. hesperus and that eye pigmentation genes are useful for tracking the successful genetic manipulation of this insect.

An agent-based model to simulate the boosted Sterile Insect Technique for fruit fly management

22556
E. G. Diouf, T. Brévault, S. Ndiaye, E. Faye, A. Chailleux, P. Diatta and C. Piou,  Ecological Modelling,  468:109951. 2022-03-18 14:58:33.
The sterile insect technique (SIT) is a method of biological control of pests and disease vector insects. It includes mass-rearing and release of sterile males of the target species so that wild females mated with sterile males would not produce offspring. An innovative version of this technique, called boosted SIT, relies on the use of sterile males as vectors of biocides to trigger an epizootic in the wild fruit fly population. We built an agent-based model to assess the feasibility of this technique and main modalities of field implementation for the control of the Oriental fruit fly, Bactrocera dorsalis, using the entomopathogenic fungi, Metarizhium anisopliae, as a biocide. The model, called BOOSTIT (BactrOcera dOrsaliS boosTed sIT), simulates the spatio-temporal population dynamics of fruit flies in three different realistic landscape contexts. The releases of infected and uninfected sterile males were simulated and allowed the transmission of the pathogen within the wild fly population as a result of interactions between individuals. A main output was the measurement of losses in mango production. Validation of the model was done by comparing the simulated population dynamics with data from field monitoring (pheromone traps) in three landscapes of the Niayes area in Senegal. The population dynamics of wild flies were then simulated in an intensive cropping and mono-mango cultivar landscape under three scenarios: (1) without the release of sterile males, (2) with the release of non-contaminated sterile males (SIT) and (3) with the release of sterile contaminated males (boosted SIT). The results showed that SIT and boosted SIT strongly reduced the density of wild flies and the amount of infested fruits. Although parameters of the pathogen transfer between individuals need to be studied more deeply, results encourage the implementation of field trials to validate the efficacy of boosted SIT to control fruit flies.

Symbionts and gene drive: two strategies to combat vector-borne disease

21029
G.-H. Wang, J. Du, C. Y. Chu, M. Madhav, G. L. Hughes and J. Champer,  Trends in Genetics,  2022-03-18 07:56:11.
Mosquitoes bring global health problems by transmitting parasites and viruses such as malaria and dengue. Unfortunately, current insecticide-based control strategies are only moderately effective because of high cost and resistance. Thus, scalable, sustainable, and cost-effective strategies are needed for mosquito-borne disease control. Symbiont-based and genome engineering-based approaches provide new tools that show promise for meeting these criteria, enabling modification or suppression approaches. Symbiotic bacteria like Wolbachia are maternally inherited and manipulate mosquito host reproduction to enhance their vertical transmission. Genome engineering-based gene drive methods, in which mosquitoes are genetically altered to spread drive alleles throughout wild populations, are also proving to be a potentially powerful approach in the laboratory. Here, we review the latest developments in both symbionts and gene drive-based methods. We describe some notable similarities, as well as distinctions and obstacles, relating to these promising technologies.

Differential viral RNA methylation contributes to pathogen blocking in Wolbachia-colonized arthropods

20977
T. Bhattacharya, L. Yan, J. M. Crawford, H. Zaher, I. L. G. Newton and R. W. Hardy,  PLoS Pathogens,  18:e1010393. 2022-03-16 07:40:04.
Arthropod endosymbiont Wolbachia pipientis is part of a global biocontrol strategy to reduce the replication of mosquito-borne RNA viruses such as alphaviruses. We previously demonstrated the importance of a host cytosine methyltransferase, DNMT2, in Drosophila and viral RNA as a cellular target during pathogen-blocking. Here we report a role for DNMT2 in Wolbachia-induced alphavirus inhibition in Aedes species. Expression of DNMT2 in mosquito tissues, including the salivary glands, is elevated upon virus infection. Notably, this is suppressed in Wolbachia-colonized animals, coincident with reduced virus replication and decreased infectivity of progeny virus. Ectopic expression of DNMT2 in cultured Aedes cells is proviral, increasing progeny virus infectivity, and this effect of DNMT2 on virus replication and infectivity is dependent on its methyltransferase activity. Finally, examining the effects of Wolbachia on modifications of viral RNA by LC-MS show a decrease in the amount of 5-methylcytosine modification consistent with the down-regulation of DNMT2 in Wolbachia colonized mosquito cells and animals. Collectively, our findings support the conclusion that disruption of 5-methylcytosine modification of viral RNA is a vital mechanism operative in pathogen blocking. These data also emphasize the essential role of epitranscriptomic modifications in regulating fundamental alphavirus replication and transmission processes.

Could species-focused suppression of Aedes aegypti, the yellow fever mosquito, and Aedes albopictus, the tiger mosquito, affect interacting predators? An evidence synthesis from the literature

20971
J. A. S. Bonds, C. M. Collins and L.-C. Gouagna,  Pest Management Science,  2022-03-16 07:20:33.
The risks of Aedes aegypti and Aedes albopictus nuisance and vector-borne diseases are rising and the adverse effects of broad-spectrum insecticide application has promoted species-specific techniques, such as sterile insect technique (SIT) and other genetic strategies, as contenders in their control operations. When specific vector suppression is proposed, potential effects on predators and wider ecosystem are some of the first stakeholder questions. These are not the only Aedes vectors of human diseases, but are those for which SIT and genetic strategies are of most interest. They vary ecologically and in habitat origin, but both have behaviourally human-adapted forms with expanding ranges. The aquatic life stages are where predation is strongest due to greater resource predictability and limited escape opportunity. These vectors' anthropic forms usually use ephemeral water bodies and man-made containers as larval habitats; predators that occur in these are mobile, opportunistic and generalist. No literature indicates that any predator depends on larvae of either species. As adults, foraging theory predicts these mosquitoes are of low profitability to predators. Energy expended hunting and consuming will mostly outweigh their energetic benefit. Moreover, as adult biomass is mobile and largely disaggregated, any predator is likely to be a generalist and opportunist. This work, which summarises much of the literature currently available on the predators of Ae. aegypti and Ae. albopictus, indicates it is highly unlikely that any predator species depends on them. Species-specific vector control to reduce nuisance and disease is thus likely to be of negligible or limited impact on non-target predators

Evaluation of Additional Drosophila suzukii Male-Only Strains Generated Through Remobilization of an FL19 Transgene

25425
A. Yamamoto, A. K. Yadav and M. J. Scott,  Frontiers in Bioengineering and Biotechnology,  10. 2022-03-15 08:27:00.
Drosophila suzukii (D. suzukii) (Matsumura, 1931; Diptera: Drosophilidae), also known as spotted wing Drosophila, is a worldwide pest of fruits with soft skins such as blueberries and cherries. Originally from Asia, D. suzukii is now present in the Americas and Europe and has become a significant economic pest. Growers largely rely on insecticides for the control of D. suzukii. Genetic strategies offer a species-specific environmentally friendly way for suppression of D. suzukii populations. We previously developed a transgenic strain of D. suzukii that produced only males on a diet that did not contain tetracycline. The strain carried a single copy of the FL19 construct on chromosome 3. Repeated releases of an excess of FL19 males led to suppression of D. suzukii populations in laboratory cage trials. Females died as a consequence of overexpression of the tetracycline transactivator (tTA) and tTA-activated expression of the head involution defective proapoptotic gene. The aim of this study was to generate additional male-only strains that carried two copies of the FL19 transgene through crossing the original line with a piggyBac jumpstarter strain. Males that carried either two chromosome 3 or a singleX-linked transgene were identified through stronger expression of the red fluorescent protein marker gene. The brighter fluorescence of the X-linked lines was likely due to dosage compensation of the red fluorescent protein gene. In total, four X-linked lines and eleven lines with two copies on chromosome 3 were obtained, of which five were further examined. All but one of the strains produced only males on a diet without tetracycline. When crossed with wild type virgin females, all of the five two copy autosomal strains examined produced only males. However, the single copy X-linked lines did not show dominant female lethality. Five of the autosomal lines were further evaluated for productivity (egg to adult) and male competition. Based on these results, the most promising lines have been selected for future population suppression experiments with strains from different geographical locations.

Modelling homing suppression gene drive in haplodiploid organisms

21016
Y. Liu and J. Champer,  bioRxiv,  2021.10.12.464047. 2022-03-15 06:38:01.
Gene drives have shown great promise for suppression of pest populations. These engineered alleles can function by a variety of mechanisms, but the most common is the CRISPR homing drive, which converts wild-type alleles to drive alleles in the germline of heterozygotes. Some potential target species are haplodiploid, in which males develop from unfertilized eggs and thus have only one copy of each chromosome. This prevents drive conversion, a substantial disadvantage compared to diploids where drive conversion can take place in both sexes. Here, we study homing suppression gene drives in haplodiploids and find that a drive targeting a female fertility gene could still be successful. However, such drives are less powerful than in diploids and suffer more from functional resistance alleles. They are substantially more vulnerable to high resistance allele formation in the embryo due to maternally deposited Cas9 and gRNA and also to somatic cleavage activity. Examining spatial models where organisms move over a continuous landscape, we find that haplodiploid suppression drives surprisingly perform nearly as well as in diploids, possibly due to their ability to spread further before inducing strong suppression. Together, these results indicate that gene drive can potentially be used to effectively suppress haplodiploid populations.Competing Interest StatementThe authors have declared no competing interest.

Combining two Genetic Sexing Strains allows sorting of non-transgenic males for Aedes genetic control

20642
C. Lutrat, M. Burckbuchler, R. P. Olmo, R. Beugnon, A. Fontaine, T. Baldet, J. Bouyer and E. Marois,  bioRxiv,  2022.03.11.483912. 2022-03-12 08:15:59.
Chemical control of the mosquito vectors Aedes albopictus and Aedes aegypti is costly, unsustainable, and increasingly ineffective due to the spread of insecticide resistance. The Sterile Insect Technique is an autocidal control tactic that represents a valuable alternative but is limited by the slow, error-prone, and wasteful sex-separation stage. Here, we present four genetic sexing strains (two for each Aedes species) based on fluorescence markers linked to the m and M sex loci, allowing the isolation of transgenic males. Furthermore, we show how combining these sexing strains allows the production of non-transgenic males. Scaling-up would allow the sorting of 100,000 neonate male larvae in under 1.5 hour with 0.01-0.1% female contamination. The resulting males present similar survival and flight ability to laboratory-reared wild-type males. By facilitating the sorting of transgenic or non-transgenic males, these Genetic Sexing Strains should enable a major upscaling in control programmes against these major vectors.Competing Interest StatementThe authors have declared no competing interest.

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