PMID- 33657402
OWN - NLM
STAT- MEDLINE
DCOM- 20210414
LR  - 20240104
IS  - 1097-4164 (Electronic)
IS  - 1097-2765 (Print)
IS  - 1097-2765 (Linking)
VI  - 81
IP  - 7
DP  - 2021 Apr 1
TI  - Genome oligopaint via local denaturation fluorescence in situ hybridization.
PG  - 1566-1577.e8
LID - S1097-2765(21)00097-6 [pii]
LID - 10.1016/j.molcel.2021.02.011 [doi]
AB  - Cas9 in complex with a programmable guide RNA targets specific double-stranded 
      DNA for cleavage. By harnessing Cas9 as a programmable loader of superhelicase to 
      genomic DNA, we report a physiological-temperature DNA fluorescence in situ 
      hybridization (FISH) method termed genome oligopaint via local denaturation 
      (GOLD) FISH. Instead of global denaturation as in conventional DNA FISH, loading 
      a superhelicase at a Cas9-generated nick allows for local DNA denaturation, 
      reducing nonspecific binding of probes and avoiding harsh treatments such as heat 
      denaturation. GOLD FISH relies on Cas9 cleaving target DNA sequences and avoids 
      the high nuclear background associated with other genome labeling methods that 
      rely on Cas9 binding. The excellent signal brightness and specificity enable us 
      to image nonrepetitive genomic DNA loci and analyze the conformational 
      differences between active and inactive X chromosomes. Finally, GOLD FISH could 
      be used for rapid identification of HER2 gene amplification in patient tissue.
CI  - Copyright (c) 2021 Elsevier Inc. All rights reserved.
FAU - Wang, Yanbo
AU  - Wang Y
AD  - Department of Biophysics and Biophysical Chemistry, Johns Hopkins University 
      School of Medicine, Baltimore, MD 21205, USA.
FAU - Cottle, Wayne Taylor
AU  - Cottle WT
AD  - Department of Biophysics and Biophysical Chemistry, Johns Hopkins University 
      School of Medicine, Baltimore, MD 21205, USA.
FAU - Wang, Haobo
AU  - Wang H
AD  - Bloomberg School of Public Health, Johns Hopkins University School of Medicine, 
      Baltimore, MD 21205, USA.
FAU - Feng, Xinyu Ashlee
AU  - Feng XA
AD  - Department of Biology, Johns Hopkins University, Baltimore, MD 21218, USA.
FAU - Mallon, John
AU  - Mallon J
AD  - Bloomberg School of Public Health, Johns Hopkins University School of Medicine, 
      Baltimore, MD 21205, USA.
FAU - Gavrilov, Momcilo
AU  - Gavrilov M
AD  - Department of Biophysics and Biophysical Chemistry, Johns Hopkins University 
      School of Medicine, Baltimore, MD 21205, USA.
FAU - Bailey, Scott
AU  - Bailey S
AD  - Department of Biophysics and Biophysical Chemistry, Johns Hopkins University 
      School of Medicine, Baltimore, MD 21205, USA; Bloomberg School of Public Health, 
      Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
FAU - Ha, Taekjip
AU  - Ha T
AD  - Department of Biophysics and Biophysical Chemistry, Johns Hopkins University 
      School of Medicine, Baltimore, MD 21205, USA; Department of Biophysics, Johns 
      Hopkins University, Baltimore, MD 21218, USA; Department of Biomedical 
      Engineering, Johns Hopkins University, Baltimore, MD 21205, USA; Howard Hughes 
      Medical Institute, Baltimore, MD 21205, USA. Electronic address: tjha@jhu.edu.
LA  - eng
GR  - T32 GM080189/GM/NIGMS NIH HHS/United States
GR  - HHMI/Howard Hughes Medical Institute/United States
GR  - R01 GM097330/GM/NIGMS NIH HHS/United States
GR  - U01 DK127432/DK/NIDDK NIH HHS/United States
GR  - T32 GM007445/GM/NIGMS NIH HHS/United States
GR  - R35 GM122569/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20210302
PL  - United States
TA  - Mol Cell
JT  - Molecular cell
JID - 9802571
RN  - 0 (RNA, Guide, CRISPR-Cas Systems)
RN  - EC 3.1.- (CRISPR-Associated Protein 9)
SB  - IM
MH  - CRISPR-Associated Protein 9/*chemistry
MH  - *CRISPR-Cas Systems
MH  - Cell Line
MH  - Female
MH  - Fibroblasts/chemistry/metabolism
MH  - *Hot Temperature
MH  - Humans
MH  - *In Situ Hybridization, Fluorescence
MH  - *Nucleic Acid Denaturation
MH  - RNA, Guide, CRISPR-Cas Systems/*chemistry
PMC - PMC8026568
MID - NIHMS1674313
OTO - NOTNLM
OT  - CRISPR-Cas9
OT  - DNA FISH
OT  - superhelicase
COIS- Declaration of interests An immediate family member of T.H. is a member of 
      advisory boards of Molecular Cell.
EDAT- 2021/03/04 06:00
MHDA- 2021/04/15 06:00
PMCR- 2022/04/01
CRDT- 2021/03/03 20:05
PHST- 2020/08/25 00:00 [received]
PHST- 2020/11/22 00:00 [revised]
PHST- 2021/02/04 00:00 [accepted]
PHST- 2021/03/04 06:00 [pubmed]
PHST- 2021/04/15 06:00 [medline]
PHST- 2021/03/03 20:05 [entrez]
PHST- 2022/04/01 00:00 [pmc-release]
AID - S1097-2765(21)00097-6 [pii]
AID - 10.1016/j.molcel.2021.02.011 [doi]
PST - ppublish
SO  - Mol Cell. 2021 Apr 1;81(7):1566-1577.e8. doi: 10.1016/j.molcel.2021.02.011. Epub 
      2021 Mar 2.