BTN1A1 Knockout UM-UC-3 Polyclonal Cells represent a genetically heterogeneous population of the UM-UC-3 human urothelial carcinoma cell line that has been subjected to CRISPR/Cas9-mediated gene disruption targeting the BTN1A1 locus. This polyclonal knockout product provides a pooled population of cells harboring diverse loss-of-function mutations in BTN1A1, enabling functional studies without the clonal selection biases inherent in single-cell?Cderived knockout lines. The use of a polyclonal format preserves a broader representation of the parental cell line??s genetic and phenotypic variability, which can be advantageous for modeling tumor heterogeneity in vitro. The knockout is generated using a ribonucleoprotein-based CRISPR/Cas9 delivery approach, resulting in efficient disruption of BTN1A1 expression across the population, as confirmed by standard protein-level and transcript-level analyses.
The host cell line, UM-UC-3, is a well-characterized model of human bladder cancer, originally established from a male patient with transitional cell carcinoma of the urinary bladder. These cells exhibit properties of malignant urothelial epithelial cells and are commonly employed in urothelial carcinoma research, including studies on tumor invasion, metastasis, and drug response. UM-UC-3 cells harbor relevant genetic alterations commonly found in bladder cancer and maintain key signaling networks associated with epithelial-to-mesenchymal transition and tumor progression. The availability of a BTN1A1 knockout in this background allows direct interrogation of butyrophilin-mediated immune modulatory mechanisms in the context of bladder cancer biology.
BTN1A1 is a member of the butyrophilin family, functioning as a negative regulator of T cell activation and participating in immune checkpoint modulation. BTN1A1 is transcriptionally upregulated by interferon-gamma and NF-??B signaling in response to inflammatory cytokines, linking it to tumor-associated immune responses. Mechanistically, BTN1A1 interacts with the T cell receptor (TCR) complex and recruits the tyrosine phosphatase SHP-1, leading to dephosphorylation of critical signaling mediators such as ZAP70 and attenuation of downstream T cell activation. This interaction suppresses T cell proliferation and reduces interleukin-2 (IL-2) production, thereby limiting effector T cell function. Beyond immune regulation, BTN1A1 also associates with xanthine oxidoreductase (XOR) in mammary epithelial cells, implicating it in milk fat globule secretion. In the immune synapse, the BTN1A1?CTCR?CSHP-1 axis represents a key suppressive loop that can be exploited by tumor cells for immune evasion.
In the UM-UC-3 bladder cancer model, BTN1A1 is hypothesized to contribute to immune evasion by inhibiting anti-tumor T cell responses within the tumor microenvironment. Disruption of BTN1A1 expression in these cells is therefore predicted to relieve T cell suppression, potentially enhancing tumor immunogenicity and susceptibility to immune-mediated clearance. This knockout model provides a powerful system to study how butyrophilins modulate adaptive immunity in bladder cancer and serves as a platform for evaluating the effect of BTN1A1 loss on tumor cell behavior, including changes in cytokine secretion, antigen presentation, and sensitivity to T cell attack.
The BTN1A1 Knockout UM-UC-3 Polyclonal Cells are ideally suited for a range of immuno-oncology applications, including functional characterization of BTN1A1 in tumor immune evasion, drug target validation, and T cell response assays. Typical experimental workflows include co-culture assays with antigen-specific T cells to measure proliferation and cytokine release (e.g., IL-2, IFN-??), flow cytometry for immune checkpoint molecule expression, Western blotting and RT-qPCR for pathway component analysis, and migration/invasion assays to assess phenotypic changes. This product serves as a robust tool for dissecting butyrophilin-mediated signaling in bladder cancer and its impact on the anti-tumor immune response. For further technical information, please contact Ascent Research.
