一个紧而可诱导的基于dCas12f的CRISPRa平台,用于可编程的体内基因激活
Hang Wan1, Deqiang Kong2, Tao Yan1
1Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, Shanghai Academy of Natural Sciences (SANS), East China Normal University, Shanghai, China.
Nature communications
|January 8, 2026
概括
一个新的基于CRISPR的基因激活系统 (HEAL) 提供了对内源基因的精确控制. 这种适用于AAV传递的紧系统,对功能基因组学和基因疗法应用具有重大前景.
科学领域:
- 分子生物学分子生物学
- 基因编辑技术的技术
- 生物技术是生物技术.
背景情况:
- 基于CRISPR的转录激活 (CRISPRa) 对功能基因组学和治疗学至关重要.
- 目前的CRISPRa系统由于Cas蛋白的大小而面临限制,阻碍了腺相关病毒 (AAV) 传递.
- 对于体内应用,需要更高效,更紧的CRISPRa系统.
研究的目的:
- 开发一种基于CRISPR的新型,高效的转录激活系统 (HEAL),可以克服AAV传递大小的限制.
- 为精确的内源基因激活设计一个紧的模块化系统.
- 在临床前模型中证明HEAL及其变体的治疗潜力.
主要方法:
- 基于dCas12f的转录激活系统 (HEAL) 的开发,使用MS2体和外套蛋白来招募交换激活器.
- HEAL的工程用于增强DNA结合,核定位和交换激活器的招募.
- 在体外和体内验证HEAL效率,并与现有的CRISPRa系统进行比较.
- 诱导变体的开发:红光诱导的OptoHEAL和小分子诱导的ChemHEAL.
- 使用AAV传递的HEAL用于急性损伤和ChemHEAL用于肥胖的临床前研究.
主要成果:
- HEAL实现了超过10万倍的内源基因激活,超过了现有的CRISPRa系统.
- OptoHEAL和ChemHEAL已经证明了远程和精确的转录控制.
- 在一只小鼠模型中,AAV传递的HEAL成功地缓解了急性损伤,通过向介质蛋白10.
- 在肥胖小鼠中,甲状腺 stromal lymphopoietin 的 ChemHEAL 中介激活导致体重减轻.
结论:
- HEAL代表了一个模块化,紧,高效的平台,用于内源性基因激活.
- 该系统显示了促进功能基因组学研究和开发新型基因疗法的巨大潜力.
- OptoHEAL和ChemHEAL为精确的治疗干预提供了增强的控制.
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