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Updated: May 7, 2025

Live-Cell Imaging of Transcriptional Activity at DNA Double-Strand Breaks
Published on: September 20, 2021
SiCLAT: simultaneous imaging of chromatin loops and active transcription in living cells
Xin Wan1, Jie Kong2, Xiaodi Hu2
1State Key Laboratory for Complex, Severe, and Rare Diseases, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, Beijing, China. xinwan@ibms.pumc.edu.cn.
Abstract:
We present SiCLAT, which introduces a dCas9-dCas13d cassette into the mouse genome. This model enables the stable expression of both dCas9 and dCas13 proteins in diverse cell populations, facilitating concurrent labeling of DNA and RNA across various cell types. Using SiCLAT, we accurately labeled chromatin loop anchor interactions and associated gene transcription during myogenic differentiation. This imaging system offers a novel means of directly observing cis-element interactions and the corresponding gene transcription in living primary cells, thus providing real-time imaging for comprehensive mechanistic investigations of dynamic enhancer-promoter or enhancer-enhancer interactions in regulating transcription activation within living cells.

