美国马萨诸塞州综合医院Joung, J. Keith小组报道了在人细胞中由CRISPR介导的增强和定向远程激活。这一研究成果发表在2021年8月5日出版的国际学术期刊《自然-方法学》上。

研究人员发现在人细胞中利用CRISPR将人工转录因子 (aTF)靶向靶基因启动子可以显著增加远程激活的有效性和可靠性。研究人员使用这一策略定向选择目标基因能够调节多个启动子的增强子，并通过将aTF靶向嵌入位于远端序列中的单核苷酸多态性来实现对人类等位基因的选择性激活。该研究结果拓宽了表观遗传编辑工具在研究和治疗中的潜在应用。

据介绍，表观遗传编辑是一种新兴的技术，它通过aTF 来调节目标基因的表达。尽管利用aTFs靶向启动子可以显著上调人类基因表达，但由aTFs调控远端转录增强子而诱导的激活则明显减弱。

附：英文原文

Title: Augmenting and directing long-range CRISPR-mediated activation in human cells

Author: Tak, Y. Esther, Horng, Joy E., Perry, Nicholas T., Schultz, Hayley T., Iyer, Sowmya, Yao, Qiuming, Zou, Luli S., Aryee, Martin J., Pinello, Luca, Joung, J. Keith

Issue&Volume: 2021-08-05

Abstract: Epigenetic editing is an emerging technology that uses artificial transcription factors (aTFs) to regulate expression of a target gene. Although human genes can be robustly upregulated by targeting aTFs to promoters, the activation induced by directing aTFs to distal transcriptional enhancers is substantially less robust and consistent. Here we show that long-range activation using CRISPR-based aTFs in human cells can be made more efficient and reliable by concurrently targeting an aTF to the target gene promoter. We used this strategy to direct target gene choice for enhancers capable of regulating more than one promoter and to achieve allele-selective activation of human genes by targeting aTFs to single-nucleotide polymorphisms embedded in distally located sequences. Our results broaden the potential applications of the epigenetic editing toolbox for research and therapeutics. Long-range CRISPR activation can be enhanced by concurrent recruitment of artificial TFs to the enhancer and promoter of a target gene. This CRISPR activation system can be employed to achieve allele-selective gene upregulation by differentially targeting single-nucleotide polymorphisms embedded in enhancers or other distally located sequences.

DOI: 10.1038/s41592-021-01224-1

Source: https://www.nature.com/articles/s41592-021-01224-1