细胞循环的调制增加了CRISPR介导的同质导向DNA修复
Guoling Li1,2, Xiaohui Yang1, Xinxin Luo1
1National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, China.
Cell & bioscience
|November 25, 2023
概括
在S和G2/M阶段增强细胞周期同步,提高了CRISPR/Cas9介导基因敲进 (KI) 的效率. 小分子和过度表达策略促进动物细胞和胚胎的同质导向修复 (HDR).
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 细胞生物学 细胞生物学
背景情况:
- 在动物细胞中,同质导向修复 (HDR) 介导的基因敲进 (KI) 是无效的.
- HDR效率与细胞周期的S和G2/M阶段有关.
- 提高这些阶段细胞比例的策略可能会改善HDR.
研究的目的:
- 研究细胞周期同步对CRISPR/Cas9介导的HDR效率的影响.
- 确定将细胞循环进展和DNA修复联系起来的分子机制.
- 为了提高动物模型中的基因编辑结果.
主要方法:
- 使用细胞循环抑制剂 (多克塞尔,伊利诺特坎,诺可达,米托米辛C) 来同步细胞.
- 在动物细胞和胚胎中评估了CRISPR/Cas9介导KI效率与各种同源捐赠者.
- 进行分子分析以识别激活的信号通路.
- 通过过度表达研究了细胞循环因子的作用.
主要成果:
- 四个小分子在动物细胞和胚胎中显著促进了CRISPR/Cas9介导的KI.
- 在S/G2/M阶段的细胞周期同步导致CDK1/CCNB1积累,激活HDR因子.
- 细胞循环因子的过度表达也提高了KI效率,与小分子效应一致.
- 确定了连接细胞周期和DNA修复的常见信号通路.
结论:
- 诱导S/G2/M相同步的小分子在动物中提高了CRISPR/Cas9 HDR效率.
- 揭示了桥梁细胞循环进展和HDR活动的分子机制.
- 为改进基因修改和基因治疗的基因编辑工具提供了基础.
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