TY - JOUR
T1 - Decrosslinking enables visualization of RNA-guided endonuclease–in situ labeling signals for DNA sequences in plant tissues
AU - Nagaki, K.
AU - Yamaji, N.
N1 - Funding Information:
We would like to thank T. Ishii (Tottori University, Japan) for technical advice. This work was partly supported by grants from the Wesco Scientific Promotion Foundation (grant no. H28-13) and the Joint Research Program of Arid Land Research Center, Tottori University (grant no. 31C2002). Seeds of N. tabacum were kindly provided by Japan Tobacco Inc.
Funding Information:
We would like to thank T. Ishii (Tottori University, Japan) for technical advice.This work was partly supported by grants from the Wesco Scientific Promotion Foundation (grant no. H28-13) and the Joint Research Program of Arid Land Research Center, Tottori University (grant no. 31C2002). Seeds of N. tabacum were kindly provided by Japan Tobacco Inc.
Publisher Copyright:
© The Author(s) 2019. Published by Oxford University Press on behalf of the Society for Experimental Biology. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
PY - 2020/3/1
Y1 - 2020/3/1
N2 - 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.
AB - 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.
KW - CRISPR/Cas9
KW - Centromere
KW - Epigenetic modifications
KW - Immunohistochemistry
KW - In situ DNA visualization
KW - RNA-guided endonuclease–in situ labeling (RGEN-ISL)
KW - Telomere
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U2 - 10.1093/jxb/erz534
DO - 10.1093/jxb/erz534
M3 - Article
C2 - 31784756
AN - SCOPUS:85082391548
SN - 0022-0957
VL - 71
SP - 1792
EP - 1800
JO - Journal of Experimental Botany
JF - Journal of Experimental Botany
IS - 6
ER -