title: Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow.
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.
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Hanhui
authors.initials: H
authors.lastname: Ma
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Li-Chun
authors.initials: LC
authors.lastname: Tu
authors.affiliation: Department of Computer Science, University of Central Florida, Orlando, Florida, USA.
authors.forename: Ardalan
authors.initials: A
authors.lastname: Naseri
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Maximiliaan
authors.initials: M
authors.lastname: Huisman
authors.affiliation: Department of Computer Science, University of Central Florida, Orlando, Florida, USA.
authors.forename: Shaojie
authors.initials: S
authors.lastname: Zhang
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: David
authors.initials: D
authors.lastname: Grunwald
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Thoru
authors.initials: T
authors.lastname: Pederson
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D000076987:CRISPR-Associated Protein 9; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004720:Endonucleases / Q000235:genetics*; D000072669:Gene Editing / Q000379:methods*; D056426:Genetic Loci / Q000235:genetics; D008856:Microscopy, Fluorescence / Q000379:methods*; D012313:RNA / Q000235:genetics*; D013194:Staining and Labeling
chemical_list: D001426:Bacterial Proteins; D012313:RNA; D000076987:CRISPR-Associated Protein 9; C000606107:Cas9 endonuclease Streptococcus pyogenes; D004720:Endonucleases