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

Pihlajamaa, Päivi; Kauko, Otto; Sahu, Biswajyoti; Kivioja, Teemu; Taipale, Jussi · Nat Biotechnol · 2023

basic_science · Level V

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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.

Medical subject headings