The APOBEC3A Knockout UM-UC-3 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal population of UM-UC-3 cells carrying a targeted disruption of the APOBEC3A gene. This genetically heterogeneous pool ablates APOBEC3A cytidine deaminase function across the cell population, serving as a robust loss-of-function tool without clonal selection. The knockout model allows interrogation of APOBEC3A-dependent processes in a human bladder carcinoma epithelial context, facilitating studies of innate immunity, viral restriction, and cancer mutagenesis mechanisms.
The parental UM-UC-3 cell line is a well-characterized human urinary bladder transitional cell carcinoma epithelial line derived from a primary bladder carcinoma of a male patient. It harbors a TP53 mutation, reflecting common genetic alterations in high-grade urothelial carcinomas. Widely employed in bladder cancer research, UM-UC-3 cells are used to examine tumor biology, drug sensitivity, and oncogenic signaling pathways. Their adherent epithelial morphology and stable growth characteristics make them suitable for a broad range of in vitro assays.
APOBEC3A encodes a potent cytidine deaminase that preferentially targets single-stranded DNA and RNA. Its expression is induced by type I interferons (IFN-??/??) through IFNAR?CJAK1?CSTAT1 signaling and the transcription factor IRF1. Once expressed, APOBEC3A interacts with replication protein A (RPA) and proliferating cell nuclear antigen (PCNA) to access ssDNA substrates, catalyzing C-to-U deamination. The resulting uracil lesions are processed by the base excision repair machinery, including uracil-DNA glycosylase UNG2, apurinic/apyrimidinic endonuclease APE1, and translesion polymerase POLH. Beyond its antiviral role, APOBEC3A is a major contributor to APOBEC signature mutations in multiple cancers.
In the UM-UC-3 bladder cancer model with TP53 deficiency, APOBEC3A-mediated mutagenesis may drive genomic instability and tumor evolution, particularly under inflammatory conditions triggered by interferon signaling. Disruption of APOBEC3A in these polyclonal cells eliminates its deaminase activity, providing a valuable isogenic platform to dissect APOBEC3A-dependent mutation patterns and DNA damage responses. This knockout model enables direct assessment of how APOBEC3A influences bladder cancer cell fitness, drug resistance, and response to genotoxic agents in a p53-mutant background.
Typical research applications include evaluating APOBEC3A-mediated C-to-T mutagenesis via targeted sequencing or whole-exome analysis, investigating interferon-induced DNA damage using ??H2AX foci formation, and examining antiviral restriction mechanisms against HIV-1 or HBV. Researchers can employ Western blotting and RT-qPCR to confirm APOBEC3A disruption, immunofluorescence to assess subcellular localization changes, and clonogenic survival or cell cycle assays to study phenotypic consequences. This polyclonal knockout tool also supports drug screening for APOBEC inhibitors and mechanistic dissection of innate immune signaling. For additional information or custom requests, please contact Ascent Research.
