Visualizing the Nucleome Using the CRISPR-Cas9 System: From in Vitro to in Vivo

Overview

Journal Biochemistry (Mosc)

Specialty Biochemistry

Date 2023 Apr 17

PMID 37069118

Authors

Liliya G Maloshenok

Gerel A Abushinova

Alexandra Yu Ryazanova

Sergey A Bruskin

Victoria V Zherdeva

Affiliations

Soon will be listed here.

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.

Citing Articles

A Comparison of Two Versions of the CRISPR-Sirius System for the Live-Cell Visualization of the Borders of Topologically Associating Domains.

Viushkov V, Lomov N, Rubtsov M Cells. 2024; 13(17.

PMID: 39273012 PMC: 11394217. DOI: 10.3390/cells13171440.

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