A Survey of Genome Editing Activity for 16 Cas12a Orthologs

© 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 en...

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Main Authors: Zetsche, Bernd, Abudayyeh, Omar O, Gootenberg, Jonathan S, Scott, David A, Zhang, Feng
Format: Article
Language:English
Published: Keio Journal of Medicine 2021
Online Access:https://hdl.handle.net/1721.1/138386
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author Zetsche, Bernd
Abudayyeh, Omar O
Gootenberg, Jonathan S
Scott, David A
Zhang, Feng
author_facet Zetsche, Bernd
Abudayyeh, Omar O
Gootenberg, Jonathan S
Scott, David A
Zhang, Feng
author_sort Zetsche, Bernd
collection MIT
description © 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.
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spelling mit-1721.1/1383862021-12-09T03:26:43Z A Survey of Genome Editing Activity for 16 Cas12a Orthologs Zetsche, Bernd Abudayyeh, Omar O Gootenberg, Jonathan S Scott, David A Zhang, Feng © 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. 2021-12-08T18:03:18Z 2021-12-08T18:03:18Z 2020 2021-12-08T17:58:18Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/138386 Zetsche, Bernd, Abudayyeh, Omar O, Gootenberg, Jonathan S, Scott, David A and Zhang, Feng. 2020. "A Survey of Genome Editing Activity for 16 Cas12a Orthologs." The Keio Journal of Medicine, 69 (3). en 10.2302/KJM.2019-0009-OA The Keio Journal of Medicine Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Keio Journal of Medicine Keio Journal of Medicine
spellingShingle Zetsche, Bernd
Abudayyeh, Omar O
Gootenberg, Jonathan S
Scott, David A
Zhang, Feng
A Survey of Genome Editing Activity for 16 Cas12a Orthologs
title A Survey of Genome Editing Activity for 16 Cas12a Orthologs
title_full A Survey of Genome Editing Activity for 16 Cas12a Orthologs
title_fullStr A Survey of Genome Editing Activity for 16 Cas12a Orthologs
title_full_unstemmed A Survey of Genome Editing Activity for 16 Cas12a Orthologs
title_short A Survey of Genome Editing Activity for 16 Cas12a Orthologs
title_sort survey of genome editing activity for 16 cas12a orthologs
url https://hdl.handle.net/1721.1/138386
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