This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population in which the HAVCR1 gene has been disrupted in the UM-UC-3 human bladder carcinoma epithelial cell line. The polyclonal format provides a heterogeneous pool of edited cells, enabling robust loss-of-function studies without the selection bottlenecks associated with monoclonal isolation. The targeted disruption abrogates HAVCR1 expression, offering a versatile model to investigate HAVCR1-dependent processes in a relevant epithelial cancer background.
UM-UC-3 cells were originally established from a male patient with transitional cell carcinoma of the bladder and are widely employed as a model for bladder cancer biology. This adherent epithelial line retains key characteristics of high-grade urothelial carcinoma, including tumorigenic potential and invasive properties. Its use as a host for HAVCR1 knockout allows researchers to dissect gene function specifically within the context of bladder cancer, complementing studies in immune cells where HAVCR1 (also known as TIM-1) is predominantly characterized.
HAVCR1 encodes a transmembrane glycoprotein that serves as a receptor for phosphatidylserine and the hepatitis A virus, while also functioning as a co-stimulatory molecule in T-cell activation. In T cells, HAVCR1 engagement by ligands such as phosphatidylserine or TIM-4 triggers downstream signaling through SRC family kinases, LCK, and ZAP70, leading to phosphorylation of LAT and PLC??1, and activation of PI3K/AKT and NF-??B pathways. These events promote T-cell proliferation and cytokine production. Upstream regulators include IL-4 and IFN-??, which modulate HAVCR1 expression during immune responses. The knockout disrupts this signaling cascade, impairing both immune regulatory functions and viral entry mechanisms.
In the bladder cancer context, the role of HAVCR1 is less defined but potentially significant, as ectopic or overexpression of immune receptors in epithelial tumors can influence tumor-immune interactions, apoptosis, and metastatic behavior. By ablating HAVCR1 in UM-UC-3 cells, researchers can examine its contribution to tumor cell-intrinsic signaling, resistance to apoptosis, and crosstalk with immune cells. The model is particularly suited to explore how HAVCR1-mediated recognition of phosphatidylserine on apoptotic cells or viral particles may impact cancer progression or viral oncolysis, given the bladder epithelium??s exposure to pathogens and inflammatory mediators.
This polyclonal knockout cell population is ideal for a spectrum of research applications including immuno-oncology, viral entry studies, and kidney injury modeling. Representative assays include T-cell proliferation assays to assess co-stimulatory requirements, viral binding and internalization assays for hepatitis A virus, and functional assays such as migration, invasion, and apoptosis to evaluate tumor cell behavior. Signaling studies by Western blotting or flow cytometry can dissect PI3K/AKT and NF-??B pathway alterations. For further details, please contact Ascent Research.
