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Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

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自然存在的CRISPR-Cas9关机

April Pawluk¹, Nadia Amrani², Yan Zhang²

¹Department of Biochemistry, University of Toronto, 1 King's College Circle, Toronto, ON M5S 1A8, Canada.

|December 17, 2016

概括

科学家发现新的抗CRISPR蛋白质可以抑制CRISPR-Cas9基因组编辑. 这些天然抑制剂为基因编辑技术的精确控制提供了必要的"关闭开关".

科学领域:

分子生物学
遗传学
微生物学

背景情况:

CRISPR-Cas9基因编辑提供了强大的工具, 但缺乏精确的控制机制.
之前已经确定了菌体编码的抗CRISPR,但仅用于I型CRISPR- Cas系统.
由于Cas9系统缺乏抗CRISPR,因此限制了条件基因组编辑的应用.

研究的目的:

发现专门抑制CRISPR-Cas9系统的新型抗CRISPR蛋白质.
描述这些新发现的抗CRISPR的抑制机制.
证明这些抗CRISPRs作为可控制的CRISPR-Cas9"关机"在真核细胞中的实用性.

主要方法:

生物信息查和功能测试以确定抗CRISPR候选物.
生物化学实验以确认抗CRISPRs与NmeCas9的直接结合.
在人类细胞中进行基于细胞的测试,以验证CRISPR-Cas9基因组编辑的抑制.

主要成果:

发现了针对Neisseria meningitidis Cas9 (NmeCas9) 的三个不同的抗CRISPR蛋白家族.
证明这些抗CRISPRs直接与NmeCas9结合.
使用这些蛋白质,成功抑制人类细胞中NmeCas9介导的基因组编辑.

结论:

已发现的抗CRISPR可对CRISPR- Cas9系统产生特定且强烈的抑制作用.
这些蛋白质作为可遗传编码的"关机",使得有条件的CRISPR-Cas9活动.
这一发现显著提高了CRISPR-Cas9基因组编辑的可控性和安全性.

关键词:

在CRISPR-Cas中其他国家脑膜炎菌抗CRISPR的方法基因组编辑菌体

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