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

Baohui Chen (), Wei Zou, Haiyue Xu, Ying Liang and Bo Huang ()
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Baohui Chen: Zhejiang University School of Medicine
Wei Zou: Zhejiang University School of Medicine
Haiyue Xu: Zhejiang University School of Medicine
Ying Liang: Zhejiang University School of Medicine
Bo Huang: University of California, San Francisco

Nature Communications, 2018, vol. 9, issue 1, 1-9

Abstract: 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.

Date: 2018
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DOI: 10.1038/s41467-018-07498-y

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