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Review
. 2023 Jan;88(Suppl 1):S123-S149.
doi: 10.1134/S0006297923140080.

Visualizing the Nucleome Using the CRISPR-Cas9 System: From in vitro to in vivo

Liliya G Maloshenok  1   2 , Gerel A Abushinova  1   2 , Alexandra Yu Ryazanova  1 , Sergey A Bruskin  1   2 , Victoria V Zherdeva  3
Affiliations
Review

Visualizing the Nucleome Using the CRISPR-Cas9 System: From in vitro to in vivo

Liliya G Maloshenok et al. Biochemistry (Mosc). 2023 Jan.

Abstract

One of the latest methods in modern molecular biology is labeling genomic loci in living cells using fluorescently labeled Cas protein. The NIH Foundation has made the mapping of the 4D nucleome (the three-dimensional nucleome on a timescale) a priority in the studies aimed to improve our understanding of chromatin organization. Fluorescent methods based on CRISPR-Cas are a significant step forward in visualization of genomic loci in living cells. This approach can be used for studying epigenetics, cell cycle, cellular response to external stimuli, rearrangements during malignant cell transformation, such as chromosomal translocations or damage, as well as for genome editing. In this review, we focused on the application of CRISPR-Cas fluorescence technologies as components of multimodal imaging methods for in vivo mapping of chromosomal loci, in particular, attribution of fluorescence signal to morphological and anatomical structures in a living organism. The review discusses the approaches to the highly sensitive, high-precision labeling of CRISPR-Cas components, delivery of genetically engineered constructs into cells and tissues, and promising methods for molecular imaging.

Keywords: 4D nucleome; CRISPR–Cas9; chromosome loci; fluorescent probes; molecular imaging.

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Conflict of interest statement

The authors declare no conflicts of interest. This article does not contain description of studies with the involvement of humans or animal subjects performed by any of the authors.

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