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在人体细胞中进行转录激活和基编辑的工程最小I型CRISPR-Cas系统

Jing Guo^1,2, Luyao Gong³, Haiying Yu¹

¹State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.

Nature communications

|August 23, 2024

概括

科学家们设计了一个紧的CRISPR-Cas系统 (类型I-F2级联),用于在人类细胞中进行基因编辑. 这个系统与dCas9的效率相匹配或超过,并创建了一个具有广泛编辑窗口的多功能基础编辑器.

科学领域:

分子生物学分子生物学
遗传学遗传学是一个
生物技术是生物技术.

背景情况:

I型CRISPR-Cas系统在 prokaryotes 中是有效的,但由于它们的尺寸和复杂性,在eukaryotes 中应用具有挑战性.
像SpCas9这样的现有CRISPR工具在尺寸和应用范围方面存在局限性.

研究的目的:

调查紧型I-F2级联系统在真核生物应用中的潜力.
开发新的基因编辑和调节工具,基于人类细胞中的I-F2级联.

主要方法:

设计了类型I-F2级联复合体,用于人类细胞中的转录激活.
开发了一个使用I-F2级联进行精确DNA修改的基础编辑器.
与现有系统相比,评估了工程I-F2工具的效率和编辑窗口.

主要成果:

经过工程设计的I-F2 Cascade在人类细胞中证明了高效的转录激活,与dCas9.9相比或优于dCas9.
使用I-F2 Cascade创建了一个新的基础编辑器,展示了大约30个核酸的广泛编辑窗口,具有双模分布.
基于I-F2 Cascade的基础编辑器扩展了针对基因选和功能序列破坏的目标站点.

结论:

紧型I型CRISPR-Cas系统,特别是I-F2级联系统,对于真核生物应用是可行的.
开发的基于I-F2 Cascade的工具提供了高效的基因调节和多功能基础编辑功能,具有广泛的编辑窗口.

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