A competitive precision CRISPR method to identify the fitness effects of transcription factor binding sites

Nat Biotechnol. 2023 Feb;41(2):197-203. doi: 10.1038/s41587-022-01444-6. Epub 2022 Sep 26.

Abstract

Here we describe a competitive genome editing method that measures the effect of mutations on molecular functions, based on precision CRISPR editing using template libraries with either the original or altered sequence, and a sequence tag, enabling direct comparison between original and mutated cells. Using the example of the MYC oncogene, we identify important transcriptional targets and show that E-box mutations at MYC target gene promoters reduce cellular fitness.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites / genetics
  • CRISPR-Cas Systems / genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics
  • Gene Editing* / methods
  • Promoter Regions, Genetic / genetics
  • Transcription Factors* / chemistry
  • Transcription Factors* / genetics

Substances

  • Transcription Factors