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

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

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.

MeSH terms

  • Bacterial Proteins / genetics*
  • CRISPR-Associated Protein 9
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Endonucleases / genetics*
  • Gene Editing / methods*
  • Genetic Loci / genetics
  • Microscopy, Fluorescence / methods*
  • RNA / genetics*
  • Staining and Labeling

Substances

  • Bacterial Proteins
  • RNA
  • CRISPR-Associated Protein 9
  • Cas9 endonuclease Streptococcus pyogenes
  • Endonucleases