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外核酶编辑器促进了哺乳动物细胞基因编辑的精度

Hui Shi^1,2,3, Lei Li^1,4, Shuangshuang Mu^1,4

¹China-New Zealand Joint Laboratory on Biomedicine and Health, CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Centre for Regenerative Medicine and Health, Hong Kong Institute of Science and Innovation, Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou Medical University, Guangzhou, 510530, China.

|May 20, 2024

概括

一个新的外核酶编辑器工具通过提高同质导向修复 (HDR) 效率和减少不必要的副产品来提高基因组编辑精度. 这种先进的工具为治疗基因校正提供了一种更安全,更通用的方法.

科学领域:

分子生物学分子生物学
基因编辑技术的技术
基因组学就是基因组学.

背景情况:

提高Cas9介导基因编辑的精度,包括敲进效率和副产品减少,仍然是一个重大挑战.
提高基因编辑精度的现有方法存在局限性.

研究的目的:

开发一种新的,高度精确的基因组编辑工具.
与现有的Cas9系统相比,评估新工具的效率和特异性.

主要方法:

开发基于人体外核酶1的基因组编辑工具 (外核酶编辑器).
在HDR效率,NHEJ修复频率和HDR/indel比率方面对外核酶编辑器和Cas9进行比较分析.
对外核酶编辑器与与精度增强域 (E1B,DN1S) 融合的Cas9的精度的评估.
研究NHEJ抑制的外核酶编辑器机制.
外核酶编辑器的应用用于纠正患者衍生的诱导多能干细胞.

主要成果:

与Cas9.9相比,外核酶编辑器显示了更高的HDR效率和更低的NHEJ修复频率.
外核酶编辑器实现了显著提高的HDR/indel比率,超过了Cas9和Cas9融合蛋白.
该工具在双链断裂时局部抑制了NHEJ,从而保持了基因组完整性.
使用Cas9尼克酶与外核酶编辑器进一步放大了HDR/英德尔比率.
观察到高的微同质介导端连接 (MMEJ) 效率,使得精确的大型删除.
通过使用外核酶编辑器成功纠正DMD患者衍生的iPSCs,以30.0%的HDR效率.

结论:

外核酶编辑器系统提供了一个高精度的多功能和安全的基因组编辑工具.
这项技术对治疗基因校正应用具有重大前景.

关键词:

在 DMD DMD 里面,你会发现外核酶编辑器的编辑器基因纠正基因纠正同质导向维修是同质导向维修的一种方式.基于微同质学的精确删除.

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