清脆的
Cas9
DNA
核糖核酸
基因组编辑
核酸内切酶
引导RNA
生物
回文
计算生物学
基因
核酸
反式激活crRNA
遗传学
复式(建筑)
细胞生物学
作者
Martin Jinek,Fuguo Jiang,David W. Taylor,Samuel H. Sternberg,Emine Kaya,Enbo Ma,Carolin Anders,M. Hauer,Keming Zhou,Steven Lin,Matias Kaplan,Anthony T. Iavarone,Emmanuelle Charpentier,Eva Nogales,Jennifer A. Doudna
出处
期刊:Science
[American Association for the Advancement of Science (AAAS)]
日期:2014-03-14
卷期号:343 (6176)
被引量:949
标识
DOI:10.1126/science.1247997
摘要
Type II CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems use an RNA-guided DNA endonuclease, Cas9, to generate double-strand breaks in invasive DNA during an adaptive bacterial immune response. Cas9 has been harnessed as a powerful tool for genome editing and gene regulation in many eukaryotic organisms. We report 2.6 and 2.2 angstrom resolution crystal structures of two major Cas9 enzyme subtypes, revealing the structural core shared by all Cas9 family members. The architectures of Cas9 enzymes define nucleic acid binding clefts, and single-particle electron microscopy reconstructions show that the two structural lobes harboring these clefts undergo guide RNA-induced reorientation to form a central channel where DNA substrates are bound. The observation that extensive structural rearrangements occur before target DNA duplex binding implicates guide RNA loading as a key step in Cas9 activation.
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