Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow
Department of Biochemistry and Molecular Pharmacology; RNA Therapeutics Institute
Biochemistry | Biotechnology | Cell Biology | Genomics | Molecular Biology | Molecular, Cellular, and Tissue Engineering | Structural Biology
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.
Cellular imaging, Chromosomes, Fluorescence imaging, Molecular engineering, RNA
DOI of Published Version
Nat Biotechnol. 2016 May;34(5):528-30. doi: 10.1038/nbt.3526. Epub 2016 Apr 18. Link to article on publisher's website
Ma H, Tu L, Naseri A, Huisman M, Zhang S, Grünwald D, Pederson T. (2016). Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Biochemistry and Molecular Pharmacology Publications. https://doi.org/10.1038/nbt.3526. Retrieved from https://escholarship.umassmed.edu/bmp_pp/216