A Survey of Genome Editing Activity for 16 Cas12a Orthologs

• Main Authors: Zetsche, Bernd (Author), Abudayyeh, Omar O. (Author), Gootenberg, Jonathan S (Author), Scott, David Arthur (Author), Zhang, Feng (Author)
• Other Authors: Broad Institute of MIT and Harvard (Contributor), McGovern Institute for Brain Research at MIT (Contributor), Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences (Contributor), Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Harvard University- (Contributor)
• Format: Article
• Language: English
• Published: Keio Journal of Medicine, 2022-01-06T14:32:23Z.
• Subjects: Article
• Online Access: Get fulltext

© 2019 by The Keio Journal of Medicine. The class 2 CRISPR-Cas endonuclease Cas12a (previously known as Cpf1) offers several advantages over Cas9, including the ability to process its own array and the requirement for just a single RNA guide. These attributes make Cas12a promising for many genome engineering applications. To further expand the suite of Cas12a tools available, we tested 16 Cas12a orthologs for activity in eukaryotic cells. Four of these new enzymes demonstrated targeted activity, one of which, from Moraxella bovoculi AAX11_00205 (Mb3Cas12a), exhibited robust indel formation. We also showed that Mb3Cas12a displays some tolerance for a shortened PAM (TTN versus the canonical Cas12a PAM TTTV). The addition of these enzymes to the genome editing toolbox will further expand the utility of this powerful technology.
National Institutes of Health (Grants 1R01-HG009761, 1R01-MH110049 and 1DP1-HL141201)