A competitive precision CRISPR method to identify the fitness effects of transcription factor binding sites.
Päivi PihlajamaaOtto KaukoBiswajyoti SahuTeemu KiviojaJussi TaipalePublished in: Nature biotechnology (2022)
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.
Keyphrases
- genome editing
- crispr cas
- transcription factor
- genome wide identification
- physical activity
- body composition
- induced apoptosis
- dna binding
- cell cycle arrest
- genome wide
- gene expression
- endoplasmic reticulum stress
- amino acid
- single molecule
- molecularly imprinted
- signaling pathway
- high resolution
- pi k akt
- wild type
- simultaneous determination
- genome wide analysis