腺基编辑器诱导小鼠胚胎和人类T细胞中的非目标结构变异
Leilei Wu1,2, Shutan Jiang2, Meisong Shi2
1Institute of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
Genome biology
|November 12, 2024
概括
在基因编辑中,CRISPR/Cas9和腺基编辑器 (ABE) 可能会导致意外的结构变异 (SV). 这项研究强调了对临床使用更安全,高保真基因编辑工具的需求.
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:
- 在临床使用中,CRISPR-Cas9基因编辑安全性至关重要.
- 之前的研究将Cas9与人类T细胞的无体积症联系起来.
- 来自基础编辑器的目标之外的结构变化 (SV) 是未知的.
研究的目的:
- 调查来自CRISPR/Cas9,ABE和CBE的目标之外的SV.
- 在小鼠胚胎和人类T细胞中分析SVs.
主要方法:
- 全基因组测序. 全基因组测序.
- 单细胞RNA测序. 一个单细胞RNA测序.
- 在小鼠胚胎和人类T细胞中进行分析.
主要成果:
- Cas9和ABE在小鼠胚胎中产生了目标外的SVs;CBE诱导了罕见的SVs.
- 在32.74%的具有Cas9的人类T细胞和在9.17%的具有ABE.的人类T细胞中发生了非目标的大缺失.
- Cas9诱导的形细胞激活了P53/apoptosis通路;ABE诱导的形细胞调高了细胞周期基因,并在G0阶段被阻止.
- 在其他细胞类型中观察到ABE脱效应.
- 高保真度ABE (ABE-V106W) 显著减少了目标外的SV.
结论:
- 在老鼠胚胎和人类T细胞中,CRISPR/Cas9和ABE诱导了目标外的SVs.
- 需要开发高保真基因编辑工具.
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