A Survey of Genome Editing Activity for 16 Cas12a Orthologs

Bernd Zetsche; Omar O. Abudayyeh; Jonathan S. Gootenberg; David A. Scott; Feng Zhang

doi:10.2302/kjm.2019-0009-OA

ORIGINAL ARTICLES

A Survey of Genome Editing Activity for 16 Cas12a Orthologs

Bernd Zetsche, Omar O. Abudayyeh, Jonathan S. Gootenberg, David A. Scott, Feng Zhang

Author information

Bernd Zetsche
Broad Institute of MIT and Harvard, Cambridge, USA
McGovern Institute for Brain Research at MIT, Cambridge, USA
Department of Brain and Cognitive Science, MIT, Cambridge, USA
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA
Department of Developmental Pathology, Institute of Pathology, Bonn Medical School, Bonn, Germany
Omar O. Abudayyeh
Broad Institute of MIT and Harvard, Cambridge, USA
McGovern Institute for Brain Research at MIT, Cambridge, USA
Department of Brain and Cognitive Science, MIT, Cambridge, USA
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA
Harvard-MIT Division of Health Science and Technology, Cambridge, USA
Jonathan S. Gootenberg
Broad Institute of MIT and Harvard, Cambridge, USA
McGovern Institute for Brain Research at MIT, Cambridge, USA
Department of Brain and Cognitive Science, MIT, Cambridge, USA
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA
Department of Systems Biology, Harvard Medical School, Boston, USA
David A. Scott
Broad Institute of MIT and Harvard, Cambridge, USA
McGovern Institute for Brain Research at MIT, Cambridge, USA
Department of Brain and Cognitive Science, MIT, Cambridge, USA
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA
Feng Zhang
Broad Institute of MIT and Harvard, Cambridge, USA
McGovern Institute for Brain Research at MIT, Cambridge, USA
Department of Brain and Cognitive Science, MIT, Cambridge, USA
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA

Keywords: genome editing, CRISPR-Cas, gene therapy

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Supplementary material

2020 Volume 69 Issue 3 Pages 59-65

DOI https://doi.org/10.2302/kjm.2019-0009-OA

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Abstract

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.

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