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. 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 .