A whole-genome CRISPRa screening metformin resistance related gene in prostate cancer

Metformin is a classic type II diabetes drug which has possessed anti-tumor properties for various cancers. However, different cancers do not respond to metformin with the same effectiveness or acquire resistance. Thus, searching for vulnerabilities of metformin-resistant prostate cancer is a promising strategy to improve the therapeutic efficiency. A genome-scale CRISPR-Cas9 activation library targeting 23430 genes is conducted to identify the genes that confer resistance to metformin in prostate cancer cells. Overall design: Examination of the genes that confer resistance to metformin in DU145 cells 22.5 µg of the human CRISPR/Cas9 SAM pooled library was transfected into 7.5×10^6 HEK293 T cells for 24 h. Viral supernatant was harvested, and DU145 cells were transduced at a calculated multiplicity of infection (MOI) of 0.4 followed by selection with hygromycin B (YEASEN, 60225ES03) and blasticidin (MDBio, D0120601) for 15 days. The DU145 cells were treated three rounds with metformin (100 mM) for 24 h .