The BTN1A1 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the CAL-27 human oral squamous cell carcinoma epithelial cell line, with targeted disruption of the BTN1A1 gene. This loss-of-function model enables investigation of BTN1A1-dependent processes without single-cell cloning, preserving a heterogeneous genetic background characteristic of pooled CRISPR editing. It is designed for studies on butyrophilin-mediated immune regulation and lipid droplet biology in oral cancer.
The CAL-27 cell line originates from a human tongue squamous cell carcinoma and serves as a widely used adherent epithelial model for oral cancer research, retaining tumorigenic features and immune modulatory capacity. This knockout line maintains the parental tumorigenic background while ablating BTN1A1, allowing controlled comparative functional analyses of immune evasion and metabolic pathways.
BTN1A1, a butyrophilin family member structurally related to B7 co-regulatory molecules, functions in lipid droplet secretion and immune checkpoint regulation. Transcriptionally activated by prolactin/STAT5 and regulated by TNF-?? and IL-1??, BTN1A1 recruits xanthine oxidoreductase (XOR) to the membrane, promoting lipid droplet envelopment and secretion, and interacts with perilipin-2 at the lipid droplet surface. In immune contexts, BTN1A1 may engage inhibitory receptors on T cells, recruiting SHP-2 to attenuate T cell receptor signaling, thereby contributing to T cell co-inhibition and tumor immune evasion. This knockout disrupts these pathways, enabling dissection of BTN1A1’s dual roles.
In the CAL-27 oral squamous carcinoma background, BTN1A1 knockout provides a system to study cancer cell?Cimmune cell crosstalk in the tumor microenvironment. As oral squamous carcinomas exploit checkpoint pathways, BTN1A1 may act as a non-classical checkpoint molecule independent of PD-1/PD-L1. By comparing knockout and parental cells, researchers can assess BTN1A1’s impact on T cell suppression, cytokine production, and lipid trafficking dysregulation in cancer metabolism.
This knockout model supports functional assays including co-culture T cell suppression experiments, BODIPY staining and flow cytometry for lipid droplet quantification, immunofluorescence for subcellular localization, and RT-qPCR/Western blotting to validate downstream targets like XOR and perilipin-2. It is also suitable for high-throughput screening of BTN1A1 pathway modulators. For further technical details or custom gene-editing inquiries, contact Ascent Research.
