CRISPRoff enables spatio-temporal control of CRISPR editing

Nat Commun. 2020 Oct 7;11(1):5041. doi: 10.1038/s41467-020-18853-3.

Abstract

Following introduction of CRISPR-Cas9 components into a cell, genome editing occurs unabated until degradation of its component nucleic acids and proteins by cellular processes. This uncontrolled reaction can lead to unintended consequences including off-target editing and chromosomal translocations. To address this, we develop a method for light-induced degradation of sgRNA termed CRISPRoff. Here we show that light-induced inactivation of ribonucleoprotein attenuates genome editing within cells and allows for titratable levels of editing efficiency and spatial patterning via selective illumination.

MeSH terms

  • CRISPR-Cas Systems / genetics*
  • CRISPR-Cas Systems / radiation effects
  • Cell Line, Tumor
  • DNA Breaks, Double-Stranded
  • Feasibility Studies
  • Gene Editing / methods*
  • HEK293 Cells
  • Humans
  • Light*
  • RNA Stability / radiation effects*
  • RNA, Guide, CRISPR-Cas Systems / metabolism*
  • RNA, Guide, CRISPR-Cas Systems / radiation effects
  • Ribonucleoproteins / metabolism
  • Translocation, Genetic

Substances

  • RNA, Guide, CRISPR-Cas Systems
  • Ribonucleoproteins