The IRGQ Knockout UM-UC-3 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal cellular pool, engineered for targeted disruption of the IRGQ gene in the UM-UC-3 bladder cancer cell line. This heterogeneous population arises from a mixed pool of editing events, providing a versatile loss-of-function resource that circumvents the need for single-cell cloning. It is optimized for high-throughput functional studies of IRGQ in a physiologically relevant cellular context.
UM-UC-3 is a well-established human urinary bladder urothelial carcinoma cell line, derived from a primary transitional cell carcinoma of a male patient with a grade 4 tumor. Exhibiting an aggressive phenotype, this cell line serves as a widely adopted model for dissecting the molecular underpinnings of bladder cancer pathogenesis, including mechanisms of tumor progression and chemoresistance. The IRGQ knockout in these cells thus offers direct applicability to clinically oriented autophagy and cancer research.
IRGQ encodes an immunity-related GTPase that dynamically localizes to mitochondria and autophagosomes, where it orchestrates autophagy flux and mitochondrial homeostasis. Upon stimulation by interferon-gamma, IRGQ is transcriptionally upregulated via STAT1 and NF-??B pathways downstream of Toll-like receptor activation. Mechanistically, IRGQ interacts with ATG5, ATG12, MAP1LC3B, VDAC1, and the anti-apoptotic regulator BCL2, facilitating the formation of the ATG12-ATG5 conjugate and promoting LC3 lipidation to drive autophagosome biogenesis. It concurrently regulates VDAC1-mediated mitochondrial outer membrane permeability and reactive oxygen species (ROS) production, thereby linking autophagic degradation with cell death control. Key pathway components such as ULK1, Beclin1, VPS34, and p62/SQSTM1 function conjointly with IRGQ to modulate autophagy-dependent cellular stress responses.
In the context of urothelial carcinoma, aberrant autophagy frequently contributes to therapeutic resistance and tumor maintenance. Disruption of IRGQ in UM-UC-3 cells enables targeted interrogation of how autophagy- and mitochondria-dependent survival signals sustain bladder cancer cell fitness. This polyclonal knockout model is thus instrumental for unraveling IRGQ??s roles in tumor progression and for identifying context-specific vulnerabilities that may be exploited for therapeutic intervention.
Researchers can utilize this knockout cell pool in a broad spectrum of assays, including immunoblotting for LC3 lipidated forms and p62/SQSTM1 to monitor autophagy flux, JC-1 staining to evaluate mitochondrial membrane potential, Annexin V apoptosis analysis, MTT cell viability tests under drug treatment, as well as qPCR, co-immunoprecipitation, and immunofluorescence microscopy. The polyclonal composition ensures robust representation of diverse knockout events, facilitating reliable bulk analyses in autophagy, innate immunity, and bladder cancer biology. For additional product information or technical support, please contact Ascent Research.
