通过CRISPR-Cas12m效应器的先天可编程DNA结合,可以实现高效的基编辑
Greta Bigelyte1, Brigita Duchovska1, Rimante Zedaveinyte1
1Institute of Biotechnology, Life Sciences Center, Vilnius University, Vilnius LT-10257, Lithuania.
Nucleic acids research
|January 23, 2024
概括
新的CRISPR-Cas12m蛋白质通过DNA结合而不是裂变提供细胞原体免疫力. 这一发现使得用于基因编辑应用的紧基基编辑工具的开发成为可能.
科学领域:
- 分子生物学分子生物学
- 微生物遗传学 微生物遗传学
- 生物化学 生物化学
背景情况:
- 2类CRISPR-Cas系统,包括Cas9和Cas12核酶,通过通过RNA引导的外来DNA分裂,赋予 prokaryotic 免疫力.
- 在Cas12蛋白家族中的多样性是广泛的,目前正在进行研究以表征新型亚型及其功能.
研究的目的:
- 描述一组新型的紧型CRISPR-Cas12m (亚型V-M) 效应蛋白.
- 阐明Cas12m蛋白在赋予 prokaryotic 免疫力的作用机制.
- 探索Cas12m效应器在开发新基因编辑技术方面的潜力.
主要方法:
- 生物化学测试被用来研究Cas12m效应器的功能性质.
- 使用冷电子显微镜 (Cryo-EM) 来确定DNA结合的结构机制.
- Cas12m与腺因除氨酶 (TadA-8e) 融合,以测试其在基编辑中的有效性.
主要成果:
- 克里斯普尔-Cas12m蛋白质通过向的DNA结合提供对细菌菌体和质粒的保护,与DNA裂变不同.
- Cas12m效应器作为路障,抑制DNA转录和/或复制,触发干扰.
- 克里奥-EM结构揭示了DNA结合和干扰的分子基础,这种干扰是由Cas12m介导的.
- GoCas12m与TadA-8e的融合导致了细菌和人类细胞中高效的A-to-G基编辑.
结论:
- 这项研究扩大了对Cas12蛋白家族的理解,突出了Cas12m效应器中的DNA结合依赖干扰机制.
- 这些发现揭示了一种由DNA结合而不是分裂介导的新型细胞原体免疫机制.
- 基于Cas12m的基编辑器代表了开发紧和高效的基因编辑工具的有希望的进步.
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