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通过配对的CRISPR-Cas9尼克酶,TREX2能够有效地破坏基因组,从而产生3'-悬浮端

Yue Wang^1,2,3, Yi-Li Feng^1,2,3, Qian Liu^1,3

¹Key Laboratory of Laparoscopic Technology of Zhejiang Province, Department of General Surgery, Sir Run-Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310016, P.R. China.

Molecular therapy. Nucleic acids

|November 29, 2023

概括

TREX2外核酶增强了配对的Streptococcus pyogenes Cas9尼克酶 (SpCas9n) 在3'-悬浮端的基因组编辑效率. 将TREX2与SpCas9n融合为基因组编辑应用提供了更安全,更方便的方法.

科学领域:

分子生物学分子生物学
基因组工程是基因组工程.
生物技术是生物技术.

背景情况:

配对的Streptococcus pyogenes Cas9 尼克酶 (SpCas9n) 减少基因组编辑中的非目标效应.
在3个悬浮的DNA末端的情况下,SpCas9n的效率是有限的.

研究的目的:

通过解决3个悬浮端的局限性来增强配对SpCas9n在基因组编辑中的实用性.
调查TREX2外核酶在处理这些DNA末端中的作用.

主要方法:

测试TREX2外核酶对3个悬浮端的SpCas9n活性的影响.
分析TREX2过度表达和SpCas9n-TREX2融合对基因组破坏的影响.
执行连接分析以了解DNA末端切除.

主要成果:

宫外TREX2过度表达增加了SpCas9n基因组破坏效率,在3个悬浮端上增加了多达400倍,目标外影响最小.
在TREX2中,优先删除3个悬浮点,而不是5个悬浮点.
与自由TREX2过度表达相比,SpCas9n-TREX2融合提高了效率和安全性.

结论:

过度表达TREX2显著提高了对配SpCas9n基因组编辑效率,用于具有3个悬浮端的基底.
将TREX2与SpCas9n融合为基因组编辑提供了更方便,更安全的方法,扩大了其实际应用.

关键词:

MT:RNA/DNA编辑编辑在 SpCas9n-TREX2 聚变过程中,这就是TREX2的特点.基因组破坏基因组破坏异于目标效应的效应配对的 SpCas9 尼卡酶.

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