PMID- 29718399
OWN - NLM
STAT- MEDLINE
DCOM- 20190805
LR  - 20190805
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Print)
IS  - 0305-1048 (Linking)
VI  - 46
IP  - 13
DP  - 2018 Jul 27
TI  - A CRISPR/molecular beacon hybrid system for live-cell genomic imaging.
PG  - e80
LID - 10.1093/nar/gky304 [doi]
AB  - The clustered regularly interspersed short palindromic repeat (CRISPR) 
      gene-editing system has been repurposed for live-cell genomic imaging, but 
      existing approaches rely on fluorescent protein reporters, making sensitive and 
      continuous imaging difficult. Here, we present a fluorophore-based live-cell 
      genomic imaging system that consists of a nuclease-deactivated mutant of the Cas9 
      protein (dCas9), a molecular beacon (MB), and an engineered single-guide RNA 
      (sgRNA) harboring a unique MB target sequence (sgRNA-MTS), termed CRISPR/MB. 
      Specifically, dCas9 and sgRNA-MTS are first co-expressed to target a specific 
      locus in cells, followed by delivery of MBs that can then hybridize to MTS to 
      illuminate the target locus. We demonstrated the feasibility of this approach for 
      quantifying genomic loci, for monitoring chromatin dynamics, and for dual-color 
      imaging when using two orthogonal MB/MTS pairs. With flexibility in selecting 
      different combinations of fluorophore/quencher pairs and MB/MTS sequences, our 
      CRISPR/MB hybrid system could be a promising platform for investigating chromatin 
      activities.
FAU - Wu, Xiaotian
AU  - Wu X
AD  - Department of Biomedical Engineering, College of Engineering, Peking University, 
      Beijing 100871, China.
AD  - School of Life Sciences, Peking University, Beijing 100871, China.
FAU - Mao, Shiqi
AU  - Mao S
AD  - Department of Biomedical Engineering, College of Engineering, Peking University, 
      Beijing 100871, China.
FAU - Yang, Yantao
AU  - Yang Y
AD  - Department of Biomedical Engineering, College of Engineering, Peking University, 
      Beijing 100871, China.
FAU - Rushdi, Muaz N
AU  - Rushdi MN
AD  - Department of Biomedical Engineering, College of Engineering, Peking University, 
      Beijing 100871, China.
AD  - Wallace H Coulter Department of Biomedical Engineering, Georgia Institute of 
      Technology, Atlanta, GA 30332, USA.
FAU - Krueger, Christopher J
AU  - Krueger CJ
AD  - Department of Biomedical Engineering, College of Engineering, Peking University, 
      Beijing 100871, China.
AD  - Wallace H Coulter Department of Biomedical Engineering, Georgia Institute of 
      Technology, Atlanta, GA 30332, USA.
FAU - Chen, Antony K
AU  - Chen AK
AD  - Department of Biomedical Engineering, College of Engineering, Peking University, 
      Beijing 100871, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Chromatin)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Oligonucleotide Probes)
RN  - EC 3.1.- (CRISPR-Associated Protein 9)
MH  - CRISPR-Associated Protein 9/genetics/metabolism
MH  - *CRISPR-Cas Systems
MH  - Chromatin/metabolism
MH  - *Fluorescent Dyes
MH  - Genomics
MH  - HEK293 Cells
MH  - HeLa Cells
MH  - Humans
MH  - *Microscopy, Fluorescence
MH  - *Oligonucleotide Probes
PMC - PMC6061827
EDAT- 2018/05/03 06:00
MHDA- 2019/08/06 06:00
PMCR- 2018/04/30
CRDT- 2018/05/03 06:00
PHST- 2018/02/13 00:00 [received]
PHST- 2018/04/12 00:00 [accepted]
PHST- 2018/05/03 06:00 [pubmed]
PHST- 2019/08/06 06:00 [medline]
PHST- 2018/05/03 06:00 [entrez]
PHST- 2018/04/30 00:00 [pmc-release]
AID - 4990026 [pii]
AID - gky304 [pii]
AID - 10.1093/nar/gky304 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 2018 Jul 27;46(13):e80. doi: 10.1093/nar/gky304.