清华大学王海峰小组宣布他们的研究显示，CRISPR活细胞成像揭示了多个非重复位点的染色质动力学和增强子相互作用。相关论文发表在2025年11月6日出版的《自然—生物技术》杂志上。

研究团队介绍了CRISPR PRO-LiveFISH （Pooled gRNAs with Orthogonal bases LiveFISH）技术，该技术将扩展遗传字母技术的正交碱基与合理的单导RNA （sgRNA）设计相结合，可以有效地标记活细胞中的多个非重复基因座。优化后的方法允许同时成像多达6个基因组位点，并允许少至10个sgRNAs进行非重复位点成像，而无需信号放大。

研究组在包括原代细胞在内的多种细胞类型中演示了该方法，并将其应用于揭示增强子启动子动力学和基因组动力学与表观遗传状态之间的相关性。课题组研究人员也证明了PCDHα尽管存在空间迁移，增强子相互作用仍可能持续存在，并且BRD4维持超增强子接触，调节癌细胞中MYC癌基因的表达。CRISPR PRO-LiveFISH可以应用于活细胞中染色质动力学和基因组组织的各种研究。

据了解，现有的可视化三维基因组动态变化的方法，启动子增强子相互作用和表观遗传修饰对非重复基因座的影响是有限的。

附：英文原文

Title: CRISPR live-cell imaging reveals chromatin dynamics and enhancer interactions at multiple non-repetitive loci

Author: Liu, Meishuo, Huang, Keyun, Zhang, Jie, Li, Qingyang, Wang, Xinming, Gu, Buming, Tang, Hao, Du, Zhenhai, Hu, Liangjun, Qi, Shutao, Ma, Yu, Yu, Hongtao, Xie, Wei, Fang, Xianyang, Wang, Haifeng

Issue&Volume: 2025-11-06

Abstract: Existing methods to visualize dynamic changes in the three-dimensional genome, promoterenhancer interactions and the influence of epigenetic modifications in non-repetitive loci are limited. Here we introduce CRISPR PRO-LiveFISH (Pooled gRNAs with Orthogonal bases LiveFISH), which combines orthogonal bases from expanded genetic alphabet technology and rational single guide RNA (sgRNA) design to efficiently label multiple non-repetitive loci in living cells. The optimized method allows simultaneous imaging of up to six genomic loci and uses as few as 10 sgRNAs for non-repetitive loci imaging without signal amplification. We demonstrate the method in diverse cell types, including primary cells, and apply it to reveal enhancerpromoter dynamics and a correlation between genomic dynamics and epigenetic states. We also show that PCDHαenhancer interactions may persist despite spatial mobility and that BRD4 maintains super-enhancer contacts regulating MYC oncogene expression in cancer cells. CRISPR PRO-LiveFISH can be applied to diverse studies of chromatin dynamics and genome organization in living cells.

DOI: 10.1038/s41587-025-02887-3

Source: https://www.nature.com/articles/s41587-025-02887-3