The HAVCR1 Knockout 786-O Polyclonal Cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal population derived from the 786-O human renal cell carcinoma line, engineered to ablate expression of the HAVCR1 gene (Hepatitis A Virus Cellular Receptor 1). This loss-of-function model provides a heterogeneous pool of HAVCR1-null cells that circumvents clonal selection artifacts, making it suitable for studying gene function in a cancer-relevant epithelial background without single-cell bottleneck effects.
Host 786-O cells originate from a clear cell renal cell carcinoma (ccRCC) with a biallelic VHL mutation, leading to constitutive HIF activation and characteristic ccRCC features. As a widely used in vitro model for renal carcinoma, 786-O cells retain epithelial morphology and tumorigenic properties, enabling investigation of HAVCR1 within the context of VHL-deficient kidney cancer signaling and metabolic dysregulation.
HAVCR1 (also known as TIM-1) is a type I transmembrane glycoprotein that functions as a receptor for phosphatidylserine on apoptotic cells and for Hepatitis A virus capsid proteins. Upon ligand engagement, HAVCR1 is activated by T-cell receptor signals and cytokines such as IL-4 and TGF-??1, and it interacts with Src family kinases including Fyn and Lck. This triggers downstream phosphorylation cascades involving PI3K/AKT and ERK MAPK pathways, leading to NFAT transcription factor activation and IL-4 expression. HAVCR1 also interacts with TIM-4 and the CD3 complex to mediate phagocytosis of apoptotic debris and modulate T-cell activation and apoptosis, in part through Bcl-2 family proteins. In renal tubular epithelium, HAVCR1 upregulation during acute kidney injury promotes tubular repair and phagocytic clearance.
In the 786-O ccRCC context, HAVCR1 knockout allows dissection of its roles in tumor cell survival, migration, and response to apoptotic stimuli. Since this cell line endogenously expresses HAVCR1, disruption of the gene provides a clean background to assess its contribution to PI3K/AKT-driven proliferation, ERK-mediated migration, and regulation of Bcl-2-dependent apoptosis. Additionally, the model enables examination of viral entry mechanisms and the interplay between kidney injury repair pathways and renal cancer progression, offering a platform to identify therapeutic vulnerabilities.
Typical applications include Western blotting and RT-qPCR to confirm HAVCR1 loss; flow cytometry to quantify surface receptor absence; Annexin V-based apoptosis assays to evaluate cell death responses; phagocytosis assays to measure clearance of apoptotic cells; Hepatitis A virus binding and entry assays; and MTT-based proliferation or transwell migration assays to assess tumor cell behavior. Researchers may also employ this model in immune checkpoint modulation studies or to explore HAVCR1-dependent signaling in renal injury. For further details or to discuss custom applications, please contact Ascent Research.
