设计的循环导向RNA可以促进基于CRISPR/Cas12a和CRISPR/Cas13d的DNA和RNA编辑
Xin Zhang1,2, Xinlong Wang2, Jie Lv2
1Dongguan Institute of Clinical Cancer Research, Affiliated Dongguan Hospital, Southern Medical University, Dongguan, 523058, China.
Genome biology
|June 23, 2023
概括
循环导向RNAs (cgRNAs) 通过增加Cas12a和Cas13d系统的稳定性和效率来增强CRISPR基因编辑. 这一突破改善了DNA和RNA编辑,用于研究和潜在的基因疗法.
科学领域:
- 分子生物学分子生物学
- 基因编辑技术的技术
- 生物技术是生物技术.
背景情况:
- CRISPR/Cas12a和CRISPR/Cas13d系统是临床潜力的研究中的重要工具.
- 自由导向RNA (gRNA) 的短半衰期限制了CRISPR编辑的效率和耐用性.
- 目前的局限性阻碍了基于CRISPR的基因编辑的全面治疗应用.
研究的目的:
- 设计循环导向RNA (cgRNA),以提高gRNA的稳定性和可用性.
- 为了提高CRISPR/Cas12a和CRISPR/Cas13d系统的编辑效率和耐用性.
- 在体外和体外环境中评估cgRNAs的疗效.
主要方法:
- 循环自由导向RNAs (cgRNAs) 的工程.
- 与线性gRNA相比,测试cgRNA稳定性和Cas核酶结合.
- 通过cgRNAs评估CRISPR/Cas12a基因激活和DNA裂变效率.
- 使用cgRNAs评估CRISPR/Cas13dRNA干扰效率.
- 在小鼠肝脏模型中的体内研究.
主要成果:
- 循环gRNAs (cgRNAs) 显著提高了Cas12a编辑效率 (2.1至40.2倍) 和Cas13d RNA干扰 (1.8倍).
- 在单个和多重基因编辑应用中,cgRNAs表现出增强的性能.
- 在多个细胞系和基因组位点中保持了特异性.
- 在体内研究证实了cgRNAs在激活基因表达和在小鼠肝脏中分裂DNA方面的功效.
结论:
- 工程cgRNA为CRISPR/Cas12a和Cas13d系统提供了对线性gRNA的稳定和高效替代品.
- cgRNAs增强可编程DNA和RNA编辑,扩大了CRISPR的应用.
- 这一创新对基础研究和基因疗法的开发具有重大前景.
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