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Efficient labeling and imaging of protein-coding genes in living cells using CRISPR-Tag

Author

Listed:
  • Baohui Chen

    (Zhejiang University School of Medicine
    Zhejiang University & Zhejiang Engineering Laboratory for Stem Cell and Immunotherapy)

  • Wei Zou

    (Zhejiang University School of Medicine
    Zhejiang University)

  • Haiyue Xu

    (Zhejiang University School of Medicine)

  • Ying Liang

    (Zhejiang University School of Medicine)

  • Bo Huang

    (University of California, San Francisco
    University of California, San Francisco
    Chan Zuckerberg Biohub)

Abstract

The lack of efficient tools to image non-repetitive genes in living cells has limited our ability to explore the functional impact of the spatiotemporal dynamics of such genes. Here, we addressed this issue by developing a CRISPR-Tag system using one to four highly active sgRNAs to specifically label protein-coding genes with a high signal-to-noise ratio for visualization by wide-field fluorescence microscopy. Our approach involves assembling a CRISPR-Tag within the intron region of a fluorescent protein and then integrating this cassette to N- or C-terminus of a specific gene, which enables simultaneous real-time imaging of protein and DNA of human protein-coding genes, such as HIST2H2BE, LMNA and HSPA8 in living cells. This CRISPR-Tag system, with a minimal size of ~250 bp DNA tag, represents an easily and broadly applicable technique to study the spatiotemporal organization of genomic elements in living cells.

Suggested Citation

  • Baohui Chen & Wei Zou & Haiyue Xu & Ying Liang & Bo Huang, 2018. "Efficient labeling and imaging of protein-coding genes in living cells using CRISPR-Tag," Nature Communications, Nature, vol. 9(1), pages 1-9, December.
    • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-07498-y
    DOI: 10.1038/s41467-018-07498-y
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    Cited by:

    1. Qin Peng & Ziliang Huang & Kun Sun & Yahan Liu & Chi Woo Yoon & Reed E. S. Harrison & Danielle L. Schmitt & Linshan Zhu & Yiqian Wu & Ipek Tasan & Huimin Zhao & Jin Zhang & Sheng Zhong & Shu Chien & Y, 2022. "Engineering inducible biomolecular assemblies for genome imaging and manipulation in living cells," Nature Communications, Nature, vol. 13(1), pages 1-15, December.

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