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Nat Biotechnol. 2016 May;34(5):528-30. doi: 10.1038/nbt.3526. Epub 2016 Apr 18.

Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow.

Author information

1
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
2
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
3
Department of Computer Science, University of Central Florida, Orlando, Florida, USA.

Abstract

A lack of techniques to image multiple genomic loci in living cells has limited our ability to investigate chromosome dynamics. Here we describe CRISPRainbow, a system for labeling DNA in living cells based on nuclease-dead (d) Cas9 combined with engineered single guide RNA (sgRNA) scaffolds that bind sets of fluorescent proteins. We demonstrate simultaneous imaging of up to six chromosomal loci in individual live cells and document large differences in the dynamic properties of different chromosomal loci.

PMID:
27088723
PMCID:
PMC4864854
DOI:
10.1038/nbt.3526
[Indexed for MEDLINE]
Free PMC Article

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