(Po)STAC (Polycistronic SunTAg modified CRISPR) enables live-cell and fixed-cell super-resolution imaging of multiple genes

Nucleic Acids Res. 2018 Mar 16;46(5):e30. doi: 10.1093/nar/gkx1271.

Abstract

CRISPR/dCas9-based labeling has allowed direct visualization of genomic regions in living cells. However, poor labeling efficiency and signal-to-background ratio have limited its application to visualize genome organization using super-resolution microscopy. We developed (Po)STAC (Polycistronic SunTAg modified CRISPR) by combining CRISPR/dCas9 with SunTag labeling and polycistronic vectors. (Po)STAC enhances both labeling efficiency and fluorescence signal detected from labeled loci enabling live cell imaging as well as super-resolution fixed-cell imaging of multiple genes with high spatiotemporal resolution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics*
  • Cell Line
  • Cells, Cultured
  • Genes / genetics*
  • Genetic Vectors / genetics*
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • In Situ Hybridization, Fluorescence / methods
  • Luminescent Measurements / methods*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mice
  • Reproducibility of Results
  • Telomere / genetics
  • Telomere / metabolism
  • Time-Lapse Imaging / methods*

Substances

  • Luminescent Proteins