Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system.
Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system.
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DOI：
10.1016/j.cell.2015.09.038
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发表时间：
2015-10-22
期刊：
影响因子：
64.5
通讯作者：
Zhang F
中科院分区：
文献类型：
--
作者：
Zetsche B;Gootenberg JS;Abudayyeh OO;Slaymaker IM;Makarova KS;Essletzbichler P;Volz SE;Joung J;van der Oost J;Regev A;Koonin EV;Zhang F
微生物适应性免疫系统CRISPR通过两类RNA引导的核酸酶效应子介导了针对外国遗传因素的防御。 1类效应子利用多蛋白质复合物，而2类效应子依赖于单组分效应子蛋白，例如特征良好的CAS9。在这里，我们报告了CPF1的表征，这是一种推定的2类CRISPR效应子。我们证明CPF1介导了与CAS9不同的特征的鲁棒DNA干扰。 CPF1是缺乏tracrrna的单个RNA引导的内核酸酶，它利用了富含T的原始原始镜头相邻基序。此外，CPF1通过交错的DNA双链断裂裂解DNA。在16个CPF1家族蛋白中，我们从酸性球菌和lachnospileceae中鉴定出了两种候选酶，具有有效的人类细胞基因组编辑活性。确定这种干扰机制可以扩大我们对CRISPR-CAS系统的理解，并提高其基因组编辑应用。
The microbial adaptive immune system CRISPR mediates defense against foreign genetic elements through two classes of RNA-guided nuclease effectors. Class 1 effectors utilize multi-protein 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.