TY - JOUR
T1 - A competitive precision CRISPR method to identify the fitness effects of transcription factor binding sites
AU - Pihlajamaa, Päivi
AU - Kauko, Otto
AU - Sahu, Biswajyoti
AU - Kivioja, Teemu
AU - Taipale, Jussi
N1 - © 2022. The Author(s).
PY - 2023/2
Y1 - 2023/2
N2 - 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.
AB - 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.
KW - Binding Sites/genetics
KW - Clustered Regularly Interspaced Short Palindromic Repeats/genetics
KW - CRISPR-Cas Systems/genetics
KW - Gene Editing/methods
KW - Promoter Regions, Genetic/genetics
KW - Transcription Factors/chemistry
U2 - 10.1038/s41587-022-01444-6
DO - 10.1038/s41587-022-01444-6
M3 - Article
C2 - 36163549
SN - 1087-0156
VL - 41
SP - 197
EP - 203
JO - Nature Biotechnology
JF - Nature Biotechnology
IS - 2
ER -