Protein Structure & Function Programme, Structural Molecular Biology Group, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Nat Struct Mol Biol. 2017 Nov;24(11):882-892. doi: 10.1038/nsmb.3486. Epub 2017 Oct 16.
CRISPR-Cas is a bacterial defense system against phage infection and nucleic acid invasion. Class 2 type II CRISPR-Cas9 has also been widely used for genome engineering. Here, we review novel insights into the CRISPR class 2 type V enzymes, specifically Cpf1 and C2c1, which display different DNA-recognition and cleavage characteristics than those of Cas9, the best-characterized member of class 2. Recent structures of these ribonucleoprotein complexes that capture several stages of the endonuclease reaction have provided molecular details of recognition, unzipping and cleavage of the target DNA, allowing their comparison with Cas9. A detailed understanding of these mechanisms is crucial for improving these genome engineering tools and expanding the genomic space that can be targeted.
CRISPR-Cas 是细菌防御噬菌体感染和核酸入侵的系统。2 类 2 型 CRISPR-Cas9 也被广泛用于基因组工程。在这里，我们回顾了对 CRISPR 2 类 V 型酶，特别是 Cpf1 和 C2c1 的新见解，它们显示出与 Cas9 不同的 DNA 识别和切割特性，Cas9 是 2 类中研究得最好的成员。这些核糖核蛋白复合物的最新结构捕获了内切酶反应的几个阶段，提供了目标 DNA 识别、解链和切割的分子细节，使它们可以与 Cas9 进行比较。对这些机制的深入了解对于改进这些基因组工程工具和扩展可以靶向的基因组空间至关重要。
