Easy quantification of template-directed CRISPR/Cas9 editing

Nucleic Acids Res. 2018 Jun 1;46(10):e58. doi: 10.1093/nar/gky164.

Abstract

Template-directed CRISPR/Cas9 editing is a powerful tool for introducing subtle mutations in genomes. However, the success rate of incorporation of the desired mutations at the target site is difficult to predict and therefore must be empirically determined. Here, we adapted the widely used TIDE method for quantification of templated editing events, including point mutations. The resulting TIDER method is a rapid, cheap and accessible tool for testing and optimization of template-directed genome editing strategies. A free web tool for TIDER data analysis is available at http://tide.nki.nl.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • Cell Line
  • Embryonic Stem Cells
  • Gene Editing / methods*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • INDEL Mutation
  • Mice
  • Mutation
  • Polymerase Chain Reaction
  • RNA, Guide
  • Reproducibility of Results
  • Retinal Pigment Epithelium / cytology
  • Software*
  • Telomerase / genetics

Substances

  • RNA, Guide
  • TERT protein, human
  • Telomerase