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CATI:一种高效的基因整合方法,用于通过MMEJ抑制动物和灵长类动物胚胎

Hongyu Chen¹, Xingchen Liu^1,2, Lanxin Li³

¹Institute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science & Intelligence Technology, Chinese Academy of Sciences, 320 Yue-Yang Road, Shanghai, 200031, China.

|June 23, 2023

概括

微同质介导的末端结合 (MMEJ) 阻碍了基因编辑. 用CasRx降低聚合酶Q (Polq) 的调节可以增强同质导向修复 (HDR),从而改善基因疗法和动物模型开发.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
生物技术是生物技术.

背景情况:

同质导向修复 (HDR) 对基因疗法和创造动物模型至关重要.
微同质介导端结合 (MMEJ) 是CRISPR介导基因编辑过程中重要的DNA修复途径.
MMEJ可以与HDR竞争,降低向基因修改的效率.

研究的目的:

调查MMEJ在CRISPR中介基因编辑中的作用.
开发一种方法,通过抑制MMEJ来提高HDR效率.
评估新方法在哺乳动物胚胎中的有效性.

主要方法:

利用CasRx降低MMEJ因子聚合酶Q (Polq) 的下调.
评估了Polq下调对DNA片段和单链寡核酸 (ssODN) 的有针对性的整合的影响.
在小鼠和子胚胎中应用了CasRX辅助向整合 (CATI) 方法.

主要成果:

降低Polq的调节显著提高了小鼠胚胎中向DNA整合的效率.
CasRX辅助有针对性的集成 (CATI) 在子胚胎中的HDR效率显著改善.
对于线性化DNA片段和ssODN集成,CATI方法被证明是有效的.

结论:

在CRISPR基因编辑过程中,MMEJ是HDR的主要竞争途径.
通过CasRX调节Polq的下调是提高HDR效率的有效策略.
CATI提供了一个有前途的工具,用于生成先进的动物模型和推进基因疗法.

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