Our findings support the idea that SETD5 is identified as one of the adaptor proteins responsible for OGT substrate specificity and that SETD5-OGT-mediated O-GlcNAcylation of RNA Pol II is involved in the transcriptional induction of CSC markers and PI3K-AKT pathway-related genes, thereby contributing to the acquisition of stemness in CRC cells. This evidence concerns the gene SETD5 and colorectal carcinoma.