Journal
CELL
Volume 155, Issue 7, Pages 1479-1491Publisher
CELL PRESS
DOI: 10.1016/j.cell.2013.12.001
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Funding
- California Institute for Quantitative Biomedical Research [QB3]
- UCSF Program for Breakthroughs in Biomedical Research
- UCSF Center for Systems and Synthetic Biology
- NIH Office of The Director (OD)
- National Institute of Dental & Craniofacial Research (NIDCR)
- Boehringer Ingelheim Fonds Ph.D. fellowship
- NIH [GM081879, DA036858, GM102706, CA168370, CA096840]
- NIH Director's Early Independence Award [OD017887]
- Leukemia and Lymphoma Society
- Helen Hay Whitney Foundation
- NIH Pathway to Independence Award [GM105913]
- Howard Hughes Medical Institute
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The spatiotemporal organization and dynamics of chromatin play critical roles in regulating genome function. However, visualizing specific, endogenous genomic loci remains challenging in living cells. Here, we demonstrate such an imaging technique by repurposing the bacterial CRISPR/Cas system. Using an EGFP-tagged endonuclease-deficient Cas9 protein and a structurally optimized small guide (sg) RNA, we show robust imaging of repetitive elements in telomeres and coding genes in living cells. Furthermore, an array of sgRNAs tiling along the target locus enables the visualization of nonrepetitive genomic sequences. Using this method, we have studied telomere dynamics during elongation or disruption, the subnuclear localization of the MUC4 loci, the cohesion of replicated MUC4 loci on sister chromatids, and their dynamic behaviors during mitosis. This CRISPR imaging tool has potential to significantly improve the capacity to study the conformation and dynamics of native chromosomes in living human cells.
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