Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow.
basic_science · Level V
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- Record sourced from PubMed, PMID 27088723.
- Also identified by DOI 10.1038/nbt.3526 and PMC identifier 4864854.
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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.
Medical subject headings
- Bacterial Proteins
- Clustered Regularly Interspaced Short Palindromic Repeats
- Endonucleases
- Gene Editing
- Microscopy, Fluorescence
- RNA