Journal
ANNUAL REVIEW OF BIOPHYSICS, VOL 45
Volume 45, Issue -, Pages 1-23Publisher
ANNUAL REVIEWS
DOI: 10.1146/annurev-biophys-062215-010830
Keywords
microscopy; CRISPR; Cas9; fluorescence; chromatin; nucleus
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Funding
- NIBIB NIH HHS [R33 EB019784] Funding Source: Medline
- NIH HHS [DP2 OD008479] Funding Source: Medline
- NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING [R33EB019784] Funding Source: NIH RePORTER
- OFFICE OF THE DIRECTOR, NATIONAL INSTITUTES OF HEALTH [DP2OD008479] Funding Source: NIH RePORTER
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The three-dimensional organization of the genome plays important roles in regulating the functional output of the genome and even in the maintenance of epigenetic inheritance and genome stability. Here, we review and compare a number of newly developed methods-especially those that utilize the CRISPR (clustered regularly interspaced short palindromic repeat)-Cas9 (CRISPR-associated protein 9) system-that enable the direct visualization of specific, endogenous DNA sequences in living cells. We also discuss the practical considerations in implementing the CRISPR imaging technique to achieve sufficient signal-to-background levels, high specificity, and high labeling efficiency. These DNA labeling methods enable tracking of the copy number, localization, and movement of genomic elements, and we discuss the potential applications of these methods in understanding the searching and targeting mechanism of the Cas9-sgRNA complex, investigating chromosome organization, and visualizing genome instability and rearrangement.
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