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紧的Cas9变体的菌体辅助进化针对一个简单的NNG PAM

Tao Qi^1,2,3, Yao Wang^2,3, Yuan Yang^2,3

¹Department of Cardiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.

Nature chemical biology

|December 5, 2023

概括

研究人员设计了一种新型紧的Cas9酶SlugCas9,以扩大基因组编辑能力. 一种新的变异,SlugCas9-NNG,识别了独特的PAM序列,使得精确的体内基因编辑用于治疗应用.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
基因编辑基因编辑

背景情况:

紧的Cas9核酶对于治疗性基因组编辑至关重要.
现有的核酶由于原空间邻基因 (PAM) 要求存在局限性,限制了基因组位点向.
之前的SlugCas9开发已经承认了一个NNGG PAM.

研究的目的:

为更广泛的基因组编辑应用设计紧的Cas9核酶,并扩展PAM识别功能.
开发一种新的腺基编辑器,用于使用工程 Cas9 变体进行体内基因破坏.

主要方法:

用菌体辅助进化来设计SlugCas9以改变PAM特异性.
产生了一个具有NNG PAM识别的SlugCas9变体 (SlugCas9-NNG).
一个基于SlugCas9-NNG的腺基编辑器被构建并通过腺相关病毒 (AAV) 传递.

主要成果:

工程 SlugCas9 显示出与 SpCas9.9 相比的活性.
成功创建了一个识别NNG PAM的SlugCas9变体,大大扩大了准范围.
SlugCas9-NNG基编辑器在体内使用单个AAV向量有效地破坏了PCSK9拼接供体和受体部位.

结论:

像SlugCas9-NNG这样的工程紧Cas9核酶增强了基因组编辑的多功能性和范围.
开发的SlugCas9-NNG基因编辑器为体内基因破坏提供了一个强大的工具,推进了治疗性基因组编辑策略.

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