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...
| Main Authors: | , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
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 |
| Summary: | 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. |
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