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通过Cas13a进行RNA引导RNA裂变的分子架构
Liang Liu1, Xueyan Li2, Jun Ma3
1Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
Cell
|August 1, 2017
概括
一个强大的RNA导向核糖酶CRISPR- Cas13a酶通过向RNA结合来激活. 这项结构研究揭示了Cas13a如何分裂RNA,为RNA操纵技术铺平了道路.
科学领域:
- 分子生物学
- 生物化学
- 遗传学
背景情况:
- CRISPR-Cas13a是一种RNA导向核糖酶,具有RNA技术的潜力.
- 了解它的激活机制对于利用它的能力至关重要.
研究的目的:
- 阐明Cas13a激活和RNA裂变的结构基础.
- 调查Cas13a在标RNA结合时的构造变化.
主要方法:
- 确定与crRNA和点RNA结合的Leptotrichia buccalis (Lbu) Cas13a的晶体结构.
- LbuCas13a-crRNA复合物的冷电子显微镜 (冷EM).
主要成果:
- 在核酶叶的中央通道内,crRNA-目标RNA双重结合.
- 向RNA结合诱导了Cas13a的显著形状变化.
- 导向-目标RNA复合体的形成激活了HEPN催化域进行RNA分裂.
结论:
- Cas13a激活涉及由导向-目标RNA双重组形成引发的构造变化.
- 激活的酶可以分裂目标RNA和附带RNA.
- 这些发现揭示了VI型CRISPR-Cas系统对RNA菌体的防御机制,并为RNA操纵工具的开发提供了信息.
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