BTN2A1 Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population of HT29 human colorectal adenocarcinoma cells with targeted disruption of the BTN2A1 gene. This loss-of-function model is designed for studying butyrophilin-mediated immunoregulation in an epithelial context. The polyclonal format ensures a heterogeneous editing landscape, avoiding clonal selection bias and providing a robust cellular platform for functional assays.
The HT29 cell line originates from a primary colon adenocarcinoma of a 44-year-old female and is widely employed as a model of intestinal epithelial cells. HT29 cells form polarized monolayers, mimic in vivo epithelial features, and are extensively utilized in colorectal cancer, inflammatory bowel disease, and epithelial barrier research. Their well-characterized growth and signaling properties make them ideal for examining epithelial-immune interactions.
BTN2A1 encodes an immunoregulatory butyrophilin that forms heterodimers with BTN3A1 to present phosphoantigens, such as HMBPP, to V??9V??2 T cells. This presentation engages the V??9V??2 TCR, triggering downstream ZAP70?CNF-??B signaling and inducing T cell activation, IFN-?? and TNF-?? secretion, and target cell cytotoxicity. BTN2A1 expression is regulated by IFN-??, NOD2 signaling, and microbial stimuli, and its functional complex includes periplakin. CRISPR-mediated knockout of BTN2A1 disrupts this phosphoantigen presentation axis, impairing V??9V??2 T cell activation and anti-tumor immune responses.
In HT29 cells, BTN2A1 knockout abrogates the capacity of tumor epithelia to directly stimulate V??9V??2 T cells, replicating immune evasion phenotypes observed in colorectal cancer. This model is invaluable for dissecting epithelial-dependent innate-like T cell activation, investigating the role of butyrophilins in autoimmune disorders, and exploring mechanisms of ?æ? T cell-mediated cytotoxicity. The knockout line provides a defined genetic system to assess the contribution of BTN2A1 to anti-tumor immunity.
Applications include co-culture assays with V??9V??2 T cells, flow cytometric analysis of activation markers, ELISA-based cytokine quantification, LDH release cytotoxicity assays, and HMBPP-pulsed presentation studies. The cells also support immunofluorescence localization, western blotting, RT-qPCR, and drug screening campaigns targeting BTN2A1-BTN3A1 interactions. For technical inquiries, please contact Ascent Research.
