在人体细胞中使用CRISPR-Cas12a和圆形RNA进行首次编辑
Ronghong Liang1, Zixin He1,2, Kevin Tianmeng Zhao3
1New Cornerstone Science Laboratory, Center for Genome Editing, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
Nature biotechnology
|January 10, 2024
概括
研究人员使用Cas12a开发了新的循环RNA介导主要编辑器 (CPE),克服了CRISPR-Cas9系统的局限性. 这些CPE为多功能基因组编辑应用提供了更高的效率和富含T的目标识别.
科学领域:
- 分子生物学分子生物学
- 基因组工程是基因组工程.
- 生物技术是生物技术.
背景情况:
- 由于系统大小和特定的PAM序列要求,CRISPR-Cas9主要编辑器面临限制.
- 现有的主要编辑系统在准和复杂化功能方面受到限制.
研究的目的:
- 开发使用较小的Cas12a蛋白质进行增强基因组编辑的新型主要编辑器系统.
- 为了克服与基于Cas9的prime编辑器相关的尺寸和PAM限制.
- 探索基于Cas12a的首席编辑器的定位偏好和复杂化潜力.
主要方法:
- 在Cas12a的基础上,开发了四个循环RNA介导主要编辑器 (CPE) 系统:niCPE,nuCPE,sniCPE和snuCPE.
- 在没有积极选择的情况下,对人类细胞的编辑效率的评估.
- 创建一个单一的SniCPE系统,将所有编辑组件结合在一个单一的促销商下.
- 通过在单个圆形RNA上排列多个导向RNA来展示多重基因编辑.
主要成果:
- CPE系统对富含T的基因组区域表现出偏好,并提供多重复合的潜力.
- 基于核酶的CPE系统实现了高达10.42%的编辑效率.
- 基于酶的CPE系统 (niCPE和sniCPE) 显示了高编辑频率,分别为24.89%和40.75%.
- 使用nickase prime编辑器实现了多达四个基因的同时编辑.
结论:
- 基于Cas12a的CPE系统为基于Cas9的首席编辑器提供了更具多样性和效率的替代方案.
- 开发的CPE系统克服了尺寸和PAM限制,使得能够针对富含T的地区.
- 多重复合的潜力和单组件系统 (one-sniCPE) 的开发显著提升了基因组编辑技术.
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