Visualizing the Genome: Experimental Approaches for Live-Cell Chromatin Imaging.

Vladimir S Viushkov, Nikolai A Lomov, Mikhail A Rubtsov, Yegor S Vassetzky

Journal: Cells 2022;11(24):4086

PMID: 36552850

Abstract

Over the years, our vision of the genome has changed from a linear molecule to that of a complex 3D structure that follows specific patterns and possesses a hierarchical organization. Currently, genomics is becoming "four-dimensional": our attention is increasingly focused on the study of chromatin dynamics over time, in the fourth dimension. Recent methods for visualizing the movements of chromatin loci in living cells by targeting fluorescent proteins can be divided into two groups. The first group requires the insertion of a special sequence into the locus of interest, to which proteins that recognize the sequence are recruited (e.g., FROS and ParB-INT methods). In the methods of the second approach, "programmed" proteins are targeted to the locus of interest (i.e., systems based on CRISPR/Cas, TALE, and zinc finger proteins). In the present review, we discuss these approaches, examine their strengths and weaknesses, and identify the key scientific problems that can be studied using these methods.

Address: Department of Molecular Biology, Faculty of Biology, Lomonosov Moscow State University, 119234 Moscow, Russia.; Department of Biochemistry, Center for Industrial Technologies and Entrepreneurship, I.M. Sechenov First Moscow State Medical University (Sechenov University), 119435 Moscow, Russia.; CNRS UMR9018, Université Paris-Saclay, Gustave Roussy, 94805 Villejuif, France.; Koltzov Institute of Developmental Biology, Russian Academy of Sciences, 119334 Moscow, Russia.
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