大规模发现神经增强剂用于 cis 调节疗法
Troy A McDiarmid1,2, Nicholas F Page3,4,5, Florence M Chardon1,2
1Department of Genome Sciences, University of Washington, Seattle, WA, USA.
bioRxiv : the preprint server for biology
|November 24, 2025
概括
这项研究使用基于CRISPR的基因激活 (CRISPRa) 识别了神经发育障碍 (NDD) 基因的活跃人类神经增强剂. 这些发现为开发针对NDD的基于CRISPR的 cis调节疗法 (CRT) 提供了资源.
科学领域:
- 神经科学是一个神经科学.
- 遗传学 是一个遗传学.
- 分子生物学分子生物学
背景情况:
- 由基因哈普洛因不足引起的神经发育障碍 (NDD) 是一个重大挑战.
- 基于CRISPR的基因激活 (CRISPRa) 提供了一种治疗策略,但需要确定神经元中的功能调节元素.
- 用CRISPRa针对cis调节元件 (cCREs) 进行cis调节疗法 (CRT) 需要对NDD风险基因进行验证的增强剂.
研究的目的:
- 为了发现NDD风险基因的功能性人类神经增强剂.
- 为了确定CRISPRa指导RNAs (gRNAs),这些指导RNAs专门调节NDD基因.
- 为开发基于CRISPRa的NDD治疗方法建立一个资源.
主要方法:
- 利用大规模并行报告测试 (MPRA) 来测试5425个候选神经元增强剂.
- 使用多重单细胞CRISPRa选与15,643个gRNA测试增强剂和促进剂在内源性环境.
- 在患者细胞系,大脑器官和人性化的小鼠模型中验证了CRISPRa gRNA.
主要成果:
- 从5,425名测试候选人中鉴定出2,422种活性神经元增强剂.
- 发现了数百个促进器和增强器向CRISPRa的gRNA,这些gRNA可调高200个NDD基因,包括91个新的增强器基因对.
- 在相关模型中使用验证的CRISPRa gRNA,确认了关键NDD基因 (SCN2A,CHD8,CTNND2,TCF4) 的治疗上调.
结论:
- 这项研究为NDD CRT开发提供了积极的,与目标相关的人类神经增强剂和gRNA试剂的全面资源.
- 这些发现促进了对神经基因调节的理解,并为发现CRT gRNA候选人建立了可概括的策略.
- 已识别的增强剂和gRNAs在NDD和其他哈普洛缺陷障碍中具有治疗应用的巨大潜力.
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