A novel tool for monitoring endogenous human MYC transcription and translation by EGFP tag insertion at the 3’ end using CRISPR-Cas9 genome editing

Abstract

AbstractThe MYC oncogene is overexpressed in over 70% of human cancers. Since its identification, the study of MYC has led to the discovery of the various ways through which oncogenes contribute to the ability of normal cells to become malignant. However, there are many aspects of MYC biology that remain unknown or controversial in terms of its regulation, targetability and downstream control of its targets. We developed two stable cell lines expressing MYC endogenously tagged with EGFP via CRISPR/Cas9-mediated genome editing. This system allows efficient detection of transcriptional activity of MYC as well the resulting fusion protein while maintaining the gene expression profiles, growth factors-associated MYC induction and growth kinetics of the parental cells. To our knowledge, this is the first report showing endogenous monitoring of MYC expression in colorectal adenocarcinoma through an EGFP tag, thus making it an efficient tool for high-throughput approaches such as genetic and drug screens.