RRC ID 58207
Author Nagaki K, Yamaji N.
Title Decrosslinking enables visualization of RNA-guided endonuclease-in situ labeling signals for DNA sequences in plant tissues.
Journal J Exp Bot
Abstract Information about the positioning of individual loci in the nucleus and the status of epigenetic modifications at these loci in each cell contained in plant tissue increases our understanding of how cells in a tissue coordinate gene expression. To obtain such information, a less damaging method of visualizing DNA in tissue that can be used with immunohistochemistry is required. Recently, a less damaging DNA visualization method using the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/associated caspase 9) system, named RNA-guided endonuclease-in situ labeling (RGEN-ISL), was reported. This system made it possible to visualize a target DNA locus in a nucleus fixed on a glass slide with a set of simple operations, but it could not be applied to cells in plant tissues. In this work, we have developed a modified RGEN-ISL method with decrosslinking that made it possible to simultaneously detect the DNA loci and immunohistochemistry signals, including histone modification, in various types of plant tissues and species.
Volume 71(6)
Pages 1792-1800
Published 2020-3-25
DOI 10.1093/jxb/erz534
PII 5648097
PMID 31784756
PMC PMC7094073
MeSH Base Sequence CRISPR-Cas Systems DNA Endonucleases* / genetics Endonucleases* / metabolism Plants / genetics* RNA RNA, Guide, Kinetoplastida*
IF 5.908
Times Cited 0
Resource
Tomato TOMJPF00001(Micro-Tom)