PMID- 25713381
OWN - NLM
STAT- MEDLINE
DCOM- 20150511
LR  - 20240104
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 112
IP  - 10
DP  - 2015 Mar 10
TI  - Multicolor CRISPR labeling of chromosomal loci in human cells.
PG  - 3002-7
LID - 10.1073/pnas.1420024112 [doi]
AB  - The intranuclear location of genomic loci and the dynamics of these loci are 
      important parameters for understanding the spatial and temporal regulation of 
      gene expression. Recently it has proven possible to visualize endogenous genomic 
      loci in live cells by the use of transcription activator-like effectors (TALEs), 
      as well as modified versions of the bacterial immunity clustered regularly 
      interspersed short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) 
      system. Here we report the design of multicolor versions of CRISPR using 
      catalytically inactive Cas9 endonuclease (dCas9) from three bacterial orthologs. 
      Each pair of dCas9-fluorescent proteins and cognate single-guide RNAs (sgRNAs) 
      efficiently labeled several target loci in live human cells. Using pairs of 
      differently colored dCas9-sgRNAs, it was possible to determine the intranuclear 
      distance between loci on different chromosomes. In addition, the fluorescence 
      spatial resolution between two loci on the same chromosome could be determined 
      and related to the linear distance between them on the chromosome's physical map, 
      thereby permitting assessment of the DNA compaction of such regions in a live 
      cell.
FAU - Ma, Hanhui
AU  - Ma H
AD  - Program in Cell and Developmental Dynamics and Department of Biochemistry and 
      Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 
      01605; hanhui.ma@umassmed.edu thoru.pederson@umassmed.edu.
FAU - Naseri, Ardalan
AU  - Naseri A
AD  - Department of Electrical Engineering and Computer Science, University of Central 
      Florida, Orlando, FL 32816; and.
FAU - Reyes-Gutierrez, Pablo
AU  - Reyes-Gutierrez P
AD  - Program in Cell and Developmental Dynamics and Department of Biochemistry and 
      Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 
      01605;
FAU - Wolfe, Scot A
AU  - Wolfe SA
AD  - Department of Molecular, Cell and Cancer Biology, University of Massachusetts 
      Medical School, Worcester, MA 01605.
FAU - Zhang, Shaojie
AU  - Zhang S
AD  - Department of Electrical Engineering and Computer Science, University of Central 
      Florida, Orlando, FL 32816; and.
FAU - Pederson, Thoru
AU  - Pederson T
AD  - Program in Cell and Developmental Dynamics and Department of Biochemistry and 
      Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 
      01605; hanhui.ma@umassmed.edu thoru.pederson@umassmed.edu.
LA  - eng
GR  - R01 GM102515/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20150223
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
SB  - IM
MH  - Cell Line, Tumor
MH  - *Chromosome Mapping
MH  - *Chromosomes, Human
MH  - *Clustered Regularly Interspaced Short Palindromic Repeats
MH  - Humans
MH  - Microscopy, Fluorescence
PMC - PMC4364232
OTO - NOTNLM
OT  - 4D nucleome
OT  - chromosomes
OT  - pericentromeric DNA
OT  - telomeres
COIS- Conflict of interest statement: The authors are named inventors on a patent 
      application related to this work filed by the University of Massachusetts.
EDAT- 2015/02/26 06:00
MHDA- 2015/05/12 06:00
PMCR- 2015/02/23
CRDT- 2015/02/26 06:00
PHST- 2015/02/26 06:00 [entrez]
PHST- 2015/02/26 06:00 [pubmed]
PHST- 2015/05/12 06:00 [medline]
PHST- 2015/02/23 00:00 [pmc-release]
AID - 1420024112 [pii]
AID - 201420024 [pii]
AID - 10.1073/pnas.1420024112 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2015 Mar 10;112(10):3002-7. doi: 
      10.1073/pnas.1420024112. Epub 2015 Feb 23.