PMID- 37335486
OWN - NLM
STAT- MEDLINE
DCOM- 20230621
LR  - 20230622
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2672
DP  - 2023
TI  - CRISPR-FISH: A CRISPR/Cas9-Based In Situ Labeling Method.
PG  - 315-335
LID - 10.1007/978-1-0716-3226-0_20 [doi]
AB  - Fluorescence in situ hybridization (FISH) has been widely used to visualize 
      target DNA sequences in fixed chromosome samples by denaturing the dsDNA to allow 
      complementary probe hybridization, thus damaging the chromatin structure by harsh 
      treatments. To overcome this limitation, a CRISPR/Cas9-based in situ labeling 
      method was developed, termed CRISPR-FISH. This method is also known as RNA-guided 
      endonuclease-in situ labeling (RGEN-ISL). Here we present different protocols for 
      the application of CRISPR-FISH on acetic acid: ethanol or formaldehyde-fixed 
      nuclei and chromosomes as well as tissue sections for labeling repetitive 
      sequences in a range of plant species. In addition, methods on how immunostaining 
      can be combined with CRISPR-FISH are provided.
CI  - (c) 2023. The Author(s), under exclusive license to Springer Science+Business 
      Media, LLC, part of Springer Nature.
FAU - Potlapalli, Bhanu Prakash
AU  - Potlapalli BP
AD  - Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, 
      Seeland, Germany.
FAU - Ishii, Takayoshi
AU  - Ishii T
AD  - Arid Land Research Center (ALRC), Tottori University, Hamasaka, Tottori, Japan.
FAU - Nagaki, Kiyotaka
AU  - Nagaki K
AD  - Institute of Plant Science and Resources, Okayama University, Chuo, Kurashiki, 
      Japan.
FAU - Somasundaram, Saravanakumar
AU  - Somasundaram S
AD  - Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, 
      Seeland, Germany.
FAU - Houben, Andreas
AU  - Houben A
AD  - Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, 
      Seeland, Germany. houben@ipk-gatersleben.de.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - In Situ Hybridization, Fluorescence/methods
MH  - *CRISPR-Cas Systems/genetics
MH  - *Chromosomes
MH  - DNA
MH  - Repetitive Sequences, Nucleic Acid
OTO - NOTNLM
OT  - CRISPR-FISH
OT  - CRISPR/Cas9
OT  - Chromatin dynamics
OT  - Cytogenetics
OT  - Dryland
OT  - FISH
OT  - Molecular cytogenetics
OT  - Plant chromosomes
OT  - RGEN-ISL
EDAT- 2023/06/19 13:08
MHDA- 2023/06/21 06:42
CRDT- 2023/06/19 11:10
PHST- 2023/06/21 06:42 [medline]
PHST- 2023/06/19 13:08 [pubmed]
PHST- 2023/06/19 11:10 [entrez]
AID - 10.1007/978-1-0716-3226-0_20 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2023;2672:315-335. doi: 10.1007/978-1-0716-3226-0_20.