Cas9的晶体结构与指导RNA和向DNA复合在一起
Hiroshi Nishimasu1, F Ann Ran2, Patrick D Hsu2
1Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, 2-11-16 Yayoi, Bunkyo, Tokyo 113-0032, Japan; JST, PRESTO, 2-11-16 Yayoi, Bunkyo, Tokyo 113-0032, Japan.
Cell
|February 18, 2014
概括
克里斯普尔-Cas9系统使用导向RNA来准DNA. 这项研究揭示了Cas9蛋白质结构,展示了它如何结合和切割DNA,从而实现了更好的基因组编辑工具.
科学领域:
- 分子生物学分子生物学
- 结构生物学 结构生物学
- 遗传学 遗传学 是一个
背景情况:
- 与CRISPR相关的内核酶Cas9是基因组编辑的关键工具.
- 通过使用单向导RNA (sgRNAs) 来实现针对特定的基因组位置.
研究的目的:
- 为了确定高分辨率的晶体结构Streptococcus pyogenes Cas9与sgRNA和目标DNA复合.
- 为了阐明RNA引导DNA向的分子机制,由Cas9.
主要方法:
- 在2.5 Å分辨率的X射线晶体学.
- 生物化学试验分析Cas9的功能.
主要成果:
- Cas9结构呈现出双叶架构,具有明显的目标识别和核酶叶.
- sgRNA:DNA异质复合体在两个叶片的接口处的槽中结合.
- HNH和RuvC核酶域定位用于DNA裂变,C端域与PAM序列相互作用.
结论:
- 这项研究提供了对Cas9介导的DNA向和裂变的详细分子理解.
- 这种结构洞察力有助于合理设计先进的基因组编辑技术.
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