I. C. Lewis, Y. Yan and G. C. Finnigan,
Access Microbiol,
3:000301.
2022.
The discovery and adaptation of CRISPR/Cas systems within molecular biology has provided advances across biological research, agriculture and human health. Genomic manipulation through use of a CRISPR nuclease and programmed guide RNAs has become a common and widely accessible practice. The identification and introduction of new engineered variants and orthologues of Cas9 as well as alternative CRISPR systems such as the type V group have provided additional molecular options for editing. These include distinct PAM requirements, staggered DNA double-strand break formation, and the ability to multiplex guide RNAs from a single expression construct. Use of CRISPR/Cas has allowed for the construction and testing of a powerful genetic architecture known as a gene drive within eukaryotic model systems. Our previous work developed a drive within budding yeast using Streptococcus pyogenes Cas9. Here, we installed the type V Francisella novicida Cas12a (Cpf1) nuclease gene and its corresponding guide RNA to power a highly efficient artificial gene drive in diploid yeast. We examined the consequence of altering guide length or introduction of individual mutational substitutions to the crRNA sequence. Cas12a-dependent gene-drive function required a guide RNA of at least 18 bp and could not tolerate most changes within the 5′ end of the crRNA.
https://www.geneconvenevi.org/wp-content/uploads/2022/01/Access-Microbiology-2.png300300David Obrochta/wp-content/uploads/2019/10/GC-color-logo-for-header-3277-x-827-1030x260.pngDavid Obrochta2022-01-14 09:19:552022-01-14 11:50:55Analysis of a Cas12a-based gene-drive system in budding yeast