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A V Smirnov¹, A N Korablev¹, I A Serova¹

¹Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.

Vavilovskii zhurnal genetiki i selektsii

|March 27, 2025

概括

在前核微注射期间的CRISPR-Cas9裂变影响了转基因的整合. 与体外线性化相比,体内Cas9裂变导致较少的结合物,这表明Cas9结合可能会阻碍DNA结合.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
基因编辑基因编辑

背景情况:

前核微注射是产生转基因生物的一个关键技术.
CRISPR-Cas9内核酶引入了针对性的DNA双链断裂,以增强转基因的整合.
了解Cas9在整合过程中与DNA修复因子的相互作用至关重要.

研究的目的:

为了比较体内Cas9分裂与体内线性化对转基因连接和整合的影响.
分析Cas9-介导裂变后转基因末端处理和结节形成的机制.

主要方法:

基于条形码的基因系统的开发,用于分析转基因重组.
在体内Cas9 RNP复合体裂变 (RNP+实验) 与体内Cas9线性化转基因 (RNP-实验) 的比较.
下一代测序 (NGS) 对条码末端和转基因-转基因结点的分析.

主要成果:

在体内Cas9裂变 (RNP+) 产生了比体内线性化 (RNP-) 更少的阳性胚胎和结合体.
NGS揭示了显著的条形码切换,表明同源重组途径的参与.
Cas9诱导的粗末端与最小的突变重新结合,而一些结点显示了Cas9末端修剪所归因的删除.

结论:

在前核微注射过程中,与Cas9结合的DNA可能会阻碍连接.
Cas9介导的裂变和随后的修复影响了转基因整合模式.
需要对体内CRISPR-Cas裂变进行进一步的研究,以阐明它对DNA修复过程的影响.

关键词:

这就是CRISPR/Cas9的作用.DNA 条形码编码 DSB维修服务提供者这就是NHEJJ.这是一台合器.同类重组 (HR) 的方法.鼠标胚胎是什么意思不同类的末端连接.无核微注射的前核微注射转基因动物转基因动物

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