目标DNA分裂后的Cpf1内核酶R循环复合物的结构
Stefano Stella1, Pablo Alcón1, Guillermo Montoya1
1Protein Structure &Function Programme, Macromolecular Crystallography Group, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, Copenhagen 2200, Denmark.
Nature
|June 1, 2017
概括
Cpf1 (CRISPR相关内核酶) 是一个基因组编辑工具. 结构分析揭示了CPF1如何解DNA以准特定的部位,从而实现精确的基因组编辑应用.
科学领域:
- 生物化学
- 分子生物学
- 遗传学
背景情况:
- Cpf1是一种RNA引导的内核酶,具有显著的基因组编辑潜力.
- 了解其DNA向机制对于优化其使用至关重要.
研究的目的:
- 为了阐明Francisella novicida Cpf1的DNA向机制.
- 为了确定Cpf1介导的DNA裂变的结构基础.
主要方法:
- 使用X射线结晶学来确定Cpf1与三链R循环的结合结构.
- 用定位基因突变来研究关键残留物的作用.
主要成果:
- 该结构显示了DNA解,crRNA-DNA杂交形成和PAM识别.
- 特定的域和图案 (PAM交互域,循环-氨酸螺旋环) 促进了DNA解锁和道.
- 一个"隔膜"结构可以防止分裂的DNA链重新结.
结论:
- 为Cpf1的RNA引导的DNA裂变机制提出了一个统一的模型.
- 这些发现为工程改进的CPF1变体提供了洞察力.
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