UBXN4 Knockout DU145 Cell Line

The UBXN4 Knockout DU145 Cell Line is a CRISPR/Cas9-edited knockout cell line, generated through targeted disruption of the UBXN4 gene in the DU145 prostate carcinoma epithelial cell line. This loss-of-function model enables precise investigation of UBXN4-dependent pathways in a well-characterized human cancer background. By eliminating UBXN4 expression, researchers can dissect its contribution to protein quality control and its impact on prostate cancer cell biology.

DU145 is an androgen receptor-negative prostate cancer cell line derived from a brain metastasis of prostate adenocarcinoma, widely used as a model for advanced, androgen-independent prostate cancer with high invasive and metastatic potential.

UBXN4 encodes a UBX domain-containing adaptor that recruits p97/VCP ATPase to polyubiquitinated substrates, facilitating extraction and delivery to proteasome or autophagosome for degradation. This places UBXN4 at the intersection of ER-associated degradation (ERAD), autophagy, and endosomal sorting. It interacts with p97/VCP, ERAD components (Derlin, HRD1), autophagy proteins (LC3, Beclin-1), and ESCRT machinery. Upstream, UBXN4 responds to unfolded protein response (UPR) and proteotoxic stress; downstream, it directs clearance of misfolded proteins, autophagy cargo receptors like p62, and endosomal cargoes.

In DU145 cells, UBXN4 knockout disrupts ERAD and autophagy, causing accumulation of misfolded proteins and altered trafficking. Given the reliance of androgen-independent prostate cancer on stress adaptation, this model is ideal for studying how loss of the p97/VCP-UBXN4 axis affects tumor fitness, drug resistance, and sensitivity to proteasome inhibitors or ER stress inducers, potentially revealing synthetic lethal interactions.

Applications include ERAD and autophagy mechanism studies, prostate cancer biology, and p97/VCP substrate identification. Researchers can perform western blotting for LC3 and p62, immunofluorescence for p97/VCP localization, flow cytometry for apoptosis/cell cycle, proteasome activity assays, migration/invasion studies, drug sensitivity testing with proteasome inhibitors or ER stress inducers, and RNA-seq for UPR gene expression. This knockout cell line is a versatile tool for dissecting protein quality control in cancer. For additional information, please contact Ascent Research.