Live-Cell Imaging of Chromatin Condensation Dynamics by CRISPR

IScience
Yuan Xue, Murat Acar

Abstract

The spatiotemporal organization of chromatin plays central roles in cellular function. The ribosomal DNA (rDNA) chromatin undergoes dynamic structural changes during mitosis and stress. Here, we developed a CRISPR-based imaging system and tracked the condensation dynamics of rDNA chromatin in live yeast cells under glucose starvation. We found that acute glucose starvation triggers rapid condensation of rDNA. Time-lapse microscopy revealed two stages for rDNA condensation: a "primary stage," when relaxed rDNA chromatin forms higher order loops or rings, and a "secondary stage," wehen the rDNA rings further condense into compact clusters. Twisting of rDNA rings accompanies the secondary stage. The condensin complex, but not the cohesin complex, is required for efficient rDNA condensation in response to glucose starvation. Furthermore, we found that the DNA helicase Sgs1 is essential for the survival of cells expressing rDNA-bound dCas9, suggesting a role for helicases in facilitating DNA replication at dCas9-binding sites.

Citations

Oct 27, 2018·Current Genetics·Matthew Robert PaulSevinç Ercan
Jul 19, 2019·Journal of Cellular Physiology·Niaz AhmadBaohong Zhang
Aug 21, 2019·Microorganisms·Rahul Mahadev ShelakeJae-Yean Kim
Dec 15, 2019·Genes·Colleen J Lawrimore, Kerry Bloom
Jul 31, 2019·Cells·Emiliano Matos-Perdomo, Félix Machín
Mar 23, 2020·Protein & Cell·Baohui ChenWei Li
Aug 9, 2019·Plant Biotechnology Journal·Mahdi Moradpour, Siti Nor Akmar Abdulah
Aug 10, 2021·Frontiers in Plant Science·Ali RazzaqFozia Saleem
Aug 20, 2021·Critical Reviews in Biotechnology·Vikram Singh, Mukesh Jain

❮ Previous
Next ❯

Methods Mentioned

BETA
immunoprecipitation
ChIP
FACS
dissection

Related Concepts

Related Feeds

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.