OBJECTIVETo explore the role of HIF1α gene in prostate cancer cell line DU145 by knocking it out with a novel gene-editing tool CRISPR/cas9 system.

METHODSA CRISPR/cas9 system with two sgRNAs targeting exon 1 of the HIF1α gene was constructed for the knock out experiment. CCK8 assay and transwell experiment were carried out to assess the effect of the knock out on the proliferation, migration and invasiveness of DU145 cells.

RESULTSThe efficiency of gene-targeting was measured through a T7E1 assaying and sequence analysis, which confirmed that the partial knock out was successful and has led to a significant decrease in the expression of HIF1α and inhibition of cell proliferation, migration and invasiveness.

CONCLUSIONA CRISPR/cas9 system for the knock out of HIF1α has been successfully constructed, which could inhibit the proliferation and migration of DU145 cells. The system can facilitate further studies of the HIF1α gene and its roles in tumorigenesis.

CRISPR-Cas Systems ; genetics ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Gene Editing ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; physiology ; Male ; Neoplasm Invasiveness ; Prostatic Neoplasms ; pathology