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可诱导的CRISPR-Cas9选平台用于询问非增殖细胞状态

Gabriele Casagrande Raffi¹, Hendrik J Kuiken², Cor Lieftink²

¹Division of Molecular Carcinogenesis, Oncode Institute, The Netherlands Cancer Institute, Amsterdam, The Netherlands.

Nature protocols

|October 8, 2025

概括

这项研究引入了针对非增殖细胞的新CRISPR查协议. 可诱导的Cas9系统能够在衰老等状态下进行精确的基因编辑,有助于发现新的治疗点.

科学领域:

分子生物学分子生物学
遗传学遗传学是一个
癌症研究癌症研究

背景情况:

克里斯普尔屏幕是识别细胞生存,增殖,耐药性和合成致命相互作用所必需的基因的强大工具.
将功能丧失CRISPR查应用于非增殖细胞是具有挑战性的,因为编辑速度缓慢,检测导向RNA脱落的灵敏度低.

研究的目的:

在非增殖状态下进行CRISPR查的详细方案.
使用可诱导的Cas9系统,对基因编辑进行精确的时间控制.

主要方法:

开发一种可诱导的Cas9系统,用于对基因编辑进行时间控制.
使用流细胞计测量来测量编辑效率.
在衰老细胞中成功进行CRISPR选的参数的优化.

主要成果:

在非增殖细胞中展示了用于CRISPR查的强有力的协议.
成功应用了在衰老细胞中识别老化标的协议.
该协议是可访问的,可以在8-12周内完成.

结论:

开发的可诱导的CRISPR查协议克服了研究非增殖细胞的局限性.
这种方法有助于识别不同领域的治疗点,包括衰老和癌症.
该协议可适应各种非增殖状态,如干细胞分化和免疫细胞发育.

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CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...