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在体内基因组编辑,使用新的Cj4Cas9基因组编辑

Tianyi Wang^1,2, Yafei Tian², Rui Yin²

¹Center for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai Engineering Research Center of Industrial Microorganisms, Fudan University, Shanghai, China.

Communications biology

|December 30, 2025

概括

研究人员发现了Cj4Cas9,一个用于基因组编辑的紧型CRISPR-Cas9系统. 像enCj4Cas9这样的工程变体提供了增强的活性和更广泛的研究和治疗向.

科学领域:

分子生物学分子生物学
遗传学遗传学是一种遗传学.
生物技术是生物技术.

背景情况:

克里斯普尔-Cas9系统是强大的基因组编辑工具.
目前正在寻找紧而高效的CRISPR-Cas9变体.
自然的CRISPR系统为基因编辑应用提供了多样化的特性.

研究的目的:

从自然资源中识别出新的,紧的CRISPR-Cas9系统.
描述和设计这些系统以提高基因组编辑能力.
评估它们在基础研究和治疗应用中的潜力.

主要方法:

使用GFP激活试验对11个CjCas9的ortolog进行选.
在小鼠模型中的体内验证 (杂虫和肝脏).
Cj4Cas9的工程,以创建一个高活性变体 (enCj4Cas9).

主要成果:

确定了七个活跃的CjCas9核酶,其中Cj4Cas9特别紧 (985个氨基酸),具有5'-NNNGRY-3' PAM.
已经证明,Tyr基因在小鼠生殖细胞中被有效地破坏,导致白化表型.
通过AAV8输送,在小鼠肝脏中展示了Pcsk9基因的体内基因编辑,降低胆固醇水平.
通过增加核酶活性和简化N3GG PAM的工程 enCj4Cas9,扩大准范围.

结论:

Cj4Cas9是一个有前途的紧型基因组编辑工具,具有独特的PAM特异性.
像enCj4Cas9这样的工程变体显著提高了编辑效率,并扩大了准范围.
这些CRISPR-Cas9系统具有基础研究和治疗开发的潜力.

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