使用CyDENT对有机细胞和核基因组进行链偏好的基因编辑
Jiacheng Hu1, Yu Sun1,2, Boshu Li1,2
1State Key Laboratory of Plant Cell and Chromosome Engineering, Center for Genome Editing, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
Nature biotechnology
|August 28, 2023
概括
介绍CyDENT基因编辑,这是一种新的无CRISPR工具,用于精确编辑核和细胞器基因组中的DNA. 这种线程选择性编辑器提高了精度,并使线粒体和叶绿体中的新编辑可能性成为可能.
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:
- 基于转录激活器样效应器 (TALE) 的基基编辑器通常使用双链DNA除氨酶,导致由于编辑两个DNA链而导致精度降低.
- 现有的方法在准特定的DNA序列方面存在局限性,特别是在器官基因组内.
研究的目的:
- 开发一种无CRISPR,有链选择性和模块化基编辑器,用于精确编辑核和细胞器DNA.
- 为了克服双链DNA脱氨酶的精度限制.
主要方法:
- 开发了CyDENT基编辑,该系统结合了一对TALEs与FokI尼克酶,单链特异性cytidine除氨酶和外核酶.
- 产生了一个单链DNA基质,用于有针对性的除.
- 在核,线粒体和叶绿体基因组中展示了编辑.
主要成果:
- 在核,线粒体和叶绿体基因组中实现了有效的基因编辑.
- 在特定的线粒体位点表现出高的链特异性 (95%) 和编辑效率 (14%).
- 通过交换去氨酶模块,在以前无法访问的GC丰富部位展示了高达20%的线粒体基编辑.
结论:
- 赛登特基编辑为编辑核和细胞器DNA提供了一个精确的,没有CRISPR的替代方案.
- 它的模块化设计允许定制,为各种基因组应用程序创建定制编辑器.
- 该系统扩大了基础编辑的范围,特别是对于具有挑战性的线粒体目标.
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