Unknown

Dataset Information

0

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


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.

SUBMITTER: Chen B 

PROVIDER: S-EPMC6265289 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature

altmetric image

Publications

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

Chen Baohui B   Zou Wei W   Xu Haiyue H   Liang Ying Y   Huang Bo B  

Nature communications 20181129 1


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 fluorescen  ...[more]

Similar Datasets

| S-EPMC2850560 | biostudies-literature
| S-EPMC6044709 | biostudies-literature
| S-EPMC8323476 | biostudies-literature
| S-EPMC8211447 | biostudies-literature
| S-EPMC1874593 | biostudies-literature
| S-EPMC5834503 | biostudies-literature
| S-EPMC6126969 | biostudies-literature
| S-EPMC8388125 | biostudies-literature
| S-EPMC5618792 | biostudies-literature
| S-EPMC5717642 | biostudies-literature