This product consists of a CRISPR/Cas9-edited polyclonal ATAD3A knockout population in the UM-UC-3 human bladder carcinoma cell line. The polyclonal format generates a heterogeneous pool of cells harboring diverse gene-disruption events within the ATAD3A locus, delivering a robust loss-of-function model without clonal isolation. This population-level approach is well-suited for functional genomic screens and studies requiring representation of heterogeneous knockout outcomes while avoiding single-clone artifacts.
The host cell line, UM-UC-3, is a grade IV transitional cell carcinoma derived from a male patient with bladder cancer. It displays aggressive in vitro characteristics, including anchorage-independent growth and metastatic potential, making it a widely employed model for urothelial carcinoma pathobiology. The ATAD3A knockout in this background permits direct interrogation of mitochondrial gene functions within a clinically relevant bladder cancer context.
ATAD3A encodes a mitochondrial inner membrane ATPase critical for mitochondrial dynamics, cholesterol trafficking, and apoptosis control. It operates at endoplasmic reticulum?Cmitochondria contact sites, forming complexes with Mitofilin (IMMT), OPA1, DRP1, VDAC1, and steroidogenic acute regulatory protein (StAR). Upstream, ATAD3A is regulated by transcription factors NRF1 and TFAM, and in cancer, c?Myc enhances its expression. Downstream, it interacts with Bcl?2 and BAX, modulating cytochrome c release and apoptotic sensitivity, while cooperating with StAR and VDAC1 to mediate cholesterol import for mitochondrial membrane integrity and steroidogenesis.
In the UM-UC-3 setting, ATAD3A disruption impairs mitochondrial cholesterol uptake and ER?Cmitochondria tethering, resulting in altered mitochondrial morphology, reduced membrane potential, and heightened apoptosis susceptibility. This recapitulates key aspects of mitochondrial dysfunction observed in advanced bladder tumors. The polyclonal knockout provides a powerful system to study ATAD3A-dependent apoptosis resistance, Wnt/???catenin signaling via mitochondrial DNA stress, and the broader contribution of mitochondrial homeostasis to urothelial carcinoma progression.
Researchers can employ this cell population for diverse assays, including Western blotting for ATAD3A depletion, mitochondrial cholesterol quantification, Annexin V apoptosis staining, JC?1 mitochondrial membrane potential measurements, proliferation and migration/invasion tests, RNA?seq transcriptomic profiling, and co?immunoprecipitation of ATAD3A interactors. The polyclonal nature also supports pooled drug screening directed at mitochondrial vulnerabilities. For further information, please contact Ascent Research.
