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一个化学诱导的CRISPR/dCas13^FCPF平台用于精确和可编程RNA调节

Sebastian Hasselbeck^1,2, Jianhui Wang^1,2, Zhaodai Bai^3,4

¹Buchmann Institute for Molecular Life Sciences, Goethe University Frankfurt am Main, 60438 Frankfurt am Main, Germany.

Journal of medicinal chemistry

|October 22, 2025

概括

化学CRISPR/dCas13^FCPF精确地针对RNA进行拼接调节,使用较低的药物剂量并减少非目标效应. 这种新的化学遗传方法为治疗中RNA调制提供了一种可控制的方法.

科学领域:

分子生物学分子生物学
在RNA治疗方面,RNA疗法.
基因编辑基因编辑

背景情况:

替代拼接调节蛋白质多样性,但其失调与癌症和神经退行等疾病有关.
目前的小分子拼接调节器缺乏特异性,导致目标外效应.
克里斯普尔/Cas13系统提供了转录水平的向,但dCas13融合效应器可以很大,并破坏RNA稳态.

研究的目的:

通过将CRISPR向与小分子作用相结合,开发一种精确可控的RNA调制系统.
创建一个模块化平台,Chem-CRISPR/dCas13^FCPF,以提高特异性和降低剂量的有针对性的剪接调节.

主要方法:

开发了Chem-CRISPR/dCas13^FCPF,这是一个通过FCPF的π标签将小分子与dCas13交联的系统.
利用CRISPRRNA (crRNA) 来引导dRfxCas13d^FCPF效应器到SMN2的第7个外显子的拼接区域.
通过测量对已知敏感转录的外子纳入和非目标效应来评估拼接调制.

主要成果:

化学CRISPR/dCas13^FCPF诱导的SMN2外因子7在配体剂量中被纳入,大约是自由RISDIPLAM的500倍.
该系统在测试中没有对其他对risdiplam敏感的转录产生可检测的非目标效应.
该平台通过crRNA重新设计显示了对其他转录的概括性.

结论:

化学CRISPR/dCas13^FCPF提供了一个近距离诱导的,化学可控制的方法,用于精确的RNA调制.
这种方法将CRISPR的准能力与剂量节约化学作用相结合,用于潜在的治疗应用.
该系统在开发有针对性的基于RNA的疗法方面取得了重大进展,其特异性提高,副作用减少.

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