Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System
The microbial adaptive immune system CRISPR mediates defense against foreign genetic elements through two classes of RNA-guided nuclease effectors. Class 1 effectors utilize multiprotein complexes, whereas Class 2 effectors rely on single-component effector proteins such as the well-characterized Ca...
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| Format: | Article |
| Language: | en_US |
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Elsevier
2016
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| Online Access: | http://hdl.handle.net/1721.1/105747 https://orcid.org/0000-0002-7979-3220 https://orcid.org/0000-0001-8794-2137 https://orcid.org/0000-0001-6656-5002 https://orcid.org/0000-0001-8567-2049 https://orcid.org/0000-0003-2782-2509 |
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| author | Makarova, Kira S. van der Oost, John Koonin, Eugene V. Zetsche, Bernd Gootenberg, Jonathan S Abudayyeh, Omar Osama Slaymaker, Ian Essletzbichler, Patrick Volz, Sara E. Joung, Julia Regev, Aviv Zhang, Feng |
| author2 | Harvard University--MIT Division of Health Sciences and Technology |
| author_facet | Harvard University--MIT Division of Health Sciences and Technology Makarova, Kira S. van der Oost, John Koonin, Eugene V. Zetsche, Bernd Gootenberg, Jonathan S Abudayyeh, Omar Osama Slaymaker, Ian Essletzbichler, Patrick Volz, Sara E. Joung, Julia Regev, Aviv Zhang, Feng |
| author_sort | Makarova, Kira S. |
| collection | MIT |
| description | The microbial adaptive immune system CRISPR mediates defense against foreign genetic elements through two classes of RNA-guided nuclease effectors. Class 1 effectors utilize multiprotein complexes, whereas Class 2 effectors rely on single-component effector proteins such as the well-characterized Cas9. Here we report characterization of Cpf1, a putative Class 2 CRISPR effector. We demonstrate that Cpf1 mediates robust DNA interference with features distinct from Cas9. Cpf1 is a single RNA-guided endonuclease lacking tracrRNA, and it utilizes a T-rich protospacer adjacent motif. Moreover, Cpf1 cleaves DNA via a staggered DNA double stranded break. Out of 16 Cpf1-family proteins, we identified two candidate enzymes, from Acidominococcus and Lachnospiraceae, with efficient genome editing activity in human cells. Identifying this mechanism of interference broadens our understanding of CRISPR-Cas systems
and advances their genome editing applications. |
| first_indexed | 2024-09-23T16:59:54Z |
| format | Article |
| id | mit-1721.1/105747 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:59:54Z |
| publishDate | 2016 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1057472022-09-29T22:59:40Z Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System Makarova, Kira S. van der Oost, John Koonin, Eugene V. Zetsche, Bernd Gootenberg, Jonathan S Abudayyeh, Omar Osama Slaymaker, Ian Essletzbichler, Patrick Volz, Sara E. Joung, Julia Regev, Aviv Zhang, Feng Harvard University--MIT Division of Health Sciences and Technology Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences McGovern Institute for Brain Research at MIT Zetsche, Bernd Gootenberg, Jonathan S Abudayyeh, Omar Osama Slaymaker, Ian Essletzbichler, Patrick Volz, Sara E. Joung, Julia Regev, Aviv Zhang, Feng The microbial adaptive immune system CRISPR mediates defense against foreign genetic elements through two classes of RNA-guided nuclease effectors. Class 1 effectors utilize multiprotein complexes, whereas Class 2 effectors rely on single-component effector proteins such as the well-characterized Cas9. Here we report characterization of Cpf1, a putative Class 2 CRISPR effector. We demonstrate that Cpf1 mediates robust DNA interference with features distinct from Cas9. Cpf1 is a single RNA-guided endonuclease lacking tracrRNA, and it utilizes a T-rich protospacer adjacent motif. Moreover, Cpf1 cleaves DNA via a staggered DNA double stranded break. Out of 16 Cpf1-family proteins, we identified two candidate enzymes, from Acidominococcus and Lachnospiraceae, with efficient genome editing activity in human cells. Identifying this mechanism of interference broadens our understanding of CRISPR-Cas systems and advances their genome editing applications. United States. Dept. of Energy (Computational Science Graduate Fellowship) National Institute of Mental Health (U.S.) (Grant 1DP1-MH100706) Poitras Foundation Vallee Foundation Simons Foundation Paul G. Allen Family Foundation New York Stem Cell Foundation Robert Metcalfe David R. Cheng 2016-12-07T21:38:01Z 2016-12-07T21:38:01Z 2015-09 2015-09 Article http://purl.org/eprint/type/JournalArticle 00928674 http://hdl.handle.net/1721.1/105747 Zetsche, Bernd et al. “Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System.” Cell 163.3 (2015): 759–771. https://orcid.org/0000-0002-7979-3220 https://orcid.org/0000-0001-8794-2137 https://orcid.org/0000-0001-6656-5002 https://orcid.org/0000-0001-8567-2049 https://orcid.org/0000-0003-2782-2509 en_US http://dx.doi.org/10.1016/j.cell.2015.09.038 Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Makarova, Kira S. van der Oost, John Koonin, Eugene V. Zetsche, Bernd Gootenberg, Jonathan S Abudayyeh, Omar Osama Slaymaker, Ian Essletzbichler, Patrick Volz, Sara E. Joung, Julia Regev, Aviv Zhang, Feng Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System |
| title | Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System |
| title_full | Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System |
| title_fullStr | Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System |
| title_full_unstemmed | Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System |
| title_short | Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System |
| title_sort | cpf1 is a single rna guided endonuclease of a class 2 crispr cas system |
| url | http://hdl.handle.net/1721.1/105747 https://orcid.org/0000-0002-7979-3220 https://orcid.org/0000-0001-8794-2137 https://orcid.org/0000-0001-6656-5002 https://orcid.org/0000-0001-8567-2049 https://orcid.org/0000-0003-2782-2509 |
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