Document Type

Article

Publication Date

4-6-2022

Publication Title

Nat Commun

Keywords

JGM, CRISPR-Cas Systems, Chromatin, Chromosomes, Clustered Regularly Interspaced Short Palindromic Repeats, Genomics, RNA, Guide

JAX Source

Nat Commun 2022 Apr 6; 13(1):1871

Volume

13

Issue

1

First Page

1871

Last Page

1871

ISSN

2041-1723

PMID

35387989

DOI

https://doi.org/10.1038/s41467-022-29343-z

Grant

HG009900, CCF-1955712, CA034196

Abstract

Three-dimensional (3D) structures of the genome are dynamic, heterogeneous and functionally important. Live cell imaging has become the leading method for chromatin dynamics tracking. However, existing CRISPR- and TALE-based genomic labeling techniques have been hampered by laborious protocols and are ineffective in labeling non-repetitive sequences. Here, we report a versatile CRISPR/Casilio-based imaging method that allows for a nonrepetitive genomic locus to be labeled using one guide RNA. We construct Casilio dual-color probes to visualize the dynamic interactions of DNA elements in single live cells in the presence or absence of the cohesin subunit RAD21. Using a three-color palette, we track the dynamic 3D locations of multiple reference points along a chromatin loop. Casilio imaging reveals intercellular heterogeneity and interallelic asynchrony in chromatin interaction dynamics, underscoring the importance of studying genome structures in 4D.

Comments

We thank the Jackson Laboratory Microscopy Service managed by Qianru Yu for
microscope usage, Erick Ratamero (Research IT Applications) for imaging analysis
support, Jane Cha (JAX Creative) for graphic design, and Research Program Development for help with manuscript editing.

This article is licensed under a Creative Commons Attribution 4.0 International License.

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