与导向RNA和目标DNA复合的Cpf1的晶体结构
Takashi Yamano1, Hiroshi Nishimasu2, Bernd Zetsche3
1Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo 113-0032, Japan.
Cell
|April 27, 2016
概括
Cpf1 (CRISPR相关蛋白) 的晶体结构揭示了它的基因组编辑机制. 这项研究提供了Cpf1如何分裂DNA的见解,有助于CRISPR-Cpf1工具的工程.
科学领域:
- 分子生物学
- 结构生物学
- 遗传学
背景情况:
- CRISPR-Cas系统是一个强大的基因组编辑工具.
- 一种与CRISPR相关的V型蛋白Cpf1是一种用于基因组编辑的多功能内核酶.
- 了解CPF1的结构和机制对于其应用至关重要.
研究的目的:
- 为了确定Acidaminococcus sp的晶体结构 Cpf1 (AsCpf1) 与指导RNA和目标DNA复合在一起.
- 阐明Cpf1的RNA引导DNA分裂活动的结构基础.
- 将AsCpf1与其他CRISPR-Cas核酶如Cas9进行比较.
主要方法:
- 在2.8 Å分辨率的X射线晶体学.
- 与同类蛋白质进行结构分析和比较.
- 生物化学分析研究DNA裂变机制.
主要成果:
- AsCpf1的晶体结构显示出中央通道中的RNA-DNA异质复合体.
- AsCpf1具有RuvC和一个用于分层DNA双链断裂的新型核酶域.
- 识别5'-TTTN-3'原空间器相邻的图案涉及底座和形状读取.
结论:
- 这项研究提供了对Cpf1介导的DNA裂变的机制性见解.
- Cpf1和Cas9之间的结构差异解释了它们不同的功能.
- 这些发现为改进的CRISPR-Cpf1基因组编辑系统奠定了基础.
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