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  3. TriTag: an integrative tool to correlate chromatin dynamics and gene expression in living cells

TriTag: an integrative tool to correlate chromatin dynamics and gene expression in living cells

  • Nucleic Acids Res. 2020 Dec 16;48(22):e127. doi: 10.1093/nar/gkaa906.
Haiyue Xu 1,  Junyan Wang 1,  Ying Liang 1,  Yujuan Fu 1,  Sihui Li 1,  Jinghan Huang 2,  Heng Xu 2 3,  Wei Zou 4 5,  Baohui Chen 1 6 7 8
Affiliations

Affiliations

  • 1 Department of Cell Biology, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China.
  • 2 School of Physics and Astronomy, Shanghai Jiao Tong University, Shanghai 200240, China.
  • 3 Institute of Natural Sciences, Shanghai Jiao Tong University, Shanghai 200240, China.
  • 4 The Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu 322000, China.
  • 5 Insititute of Translational Medicine, Zhejiang University, Hangzhou 310003, China.
  • 6 Institute of Hematology, Zhejiang University & Zhejiang Engineering Laboratory for Stem Cell and Immunotherapy, Hangzhou 310058, China.
  • 7 Zhejiang Laboratory for Systems & Precision Medicine, Zhejiang University Medical Center, Hangzhou 311121, China.
  • 8 Zhejiang Provincial Key Laboratory of Genetic & Developmental Disorders, Hangzhou 310058, China.
PMID: 33104788 DOI: 10.1093/nar/gkaa906
Abstract

A wealth of single-cell imaging studies have contributed novel insights into chromatin organization and gene regulation. However, a comprehensive understanding of spatiotemporal gene regulation requires developing tools to combine multiple monitoring systems in a single study. Here, we report a versatile tag, termed TriTag, which integrates the functional capabilities of CRISPR-Tag (DNA labeling), MS2 aptamer (RNA imaging) and Fluorescent protein (protein tracking). Using this tag, we correlate changes in chromatin dynamics with the progression of endogenous gene expression, by recording both transcriptional bursting and protein production. This strategy allows precise measurements of gene expression at single-allele resolution across the cell cycle or in response to stress. TriTag enables capturing an integrated picture of gene expression, thus providing a powerful tool to study transcriptional heterogeneity and regulation.

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