Cpf1是2类CRISPR-Cas系统中的一种单RNA引导的内切核酸酶。
Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system.
作者信息
Zetsche Bernd, Gootenberg Jonathan S, Abudayyeh Omar O, Slaymaker Ian M, Makarova Kira S, Essletzbichler Patrick, Volz Sara E, Joung Julia, van der Oost John, Regev Aviv, Koonin Eugene V, Zhang Feng
机构信息
Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Developmental Pathology, Institute of Pathology, Bonn Medical School, Sigmund Freud Street 25, 53127 Bonn, Germany.
Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
出版信息
Cell. 2015 Oct 22;163(3):759-71. doi: 10.1016/j.cell.2015.09.038. Epub 2015 Sep 25.
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 Acidaminococcus 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.
微生物适应性免疫系统CRISPR通过两类RNA引导的核酸酶效应物介导对外源遗传元件的防御。1类效应物利用多蛋白复合物，而2类效应物依赖于单组分效应蛋白，如特征明确的Cas9。在此，我们报道了一种假定的2类CRISPR效应物Cpf1的特性。我们证明Cpf1介导强大的DNA干扰，其特征不同于Cas9。Cpf1是一种缺乏反式激活crRNA的单RNA引导的内切核酸酶，它利用富含T的原间隔相邻基序。此外，Cpf1通过交错的DNA双链断裂切割DNA。在16种Cpf1家族蛋白中，我们从嗜酸菌属和毛螺菌科中鉴定出两种候选酶，它们在人类细胞中具有高效的基因组编辑活性。确定这种干扰机制拓宽了我们对CRISPR-Cas系统的理解，并推动了它们在基因组编辑中的应用。
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