CRISPR-Sirius: RNA scaffolds for signal amplification in genome imaging

Nat Methods. 2018 Nov;15(11):928-931. doi: 10.1038/s41592-018-0174-0. Epub 2018 Oct 30.

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR) guide RNA scaffolds have been adapted to carry multiple binding sites for fluorescent proteins to enhance brightness for live cell imaging of genomic loci. However, many of these modifications result in guide RNA instability and thus produce lower genome-labeling efficiency than anticipated. Here we introduce CRISPR-Sirius, based on octet arrays of aptamers conferring both enhanced guide RNA stability and brightness, and provide initial biological applications of this platform.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • A549 Cells
  • Binding Sites
  • CRISPR-Cas Systems*
  • Fluorescent Dyes / chemistry*
  • Genes, Reporter
  • Genome, Human*
  • Genomics / methods*
  • HEK293 Cells
  • Humans
  • Microscopy, Fluorescence
  • Molecular Imaging / methods*
  • RNA, Guide / genetics*

Substances

  • Fluorescent Dyes
  • RNA, Guide