BTN3A3 Knockout Jurkat Cell Line

The BTN3A3 Knockout Jurkat Cell Line is a CRISPR/Cas9-edited Jurkat T lymphoblastoid cell line with targeted disruption of the BTN3A3 gene. This knockout model provides a defined genetic background for investigating butyrophilin-mediated immune regulation. Derived from the well-characterized Jurkat T-cell leukemia line, the product is delivered as a stable, viably frozen cell line suitable for immediate culture and downstream functional assays. The gene-edited cells are produced using CRISPR/Cas9-mediated gene disruption, generating a loss-of-function model for studying phosphoantigen sensing, T-cell co-signaling, and innate-like lymphocyte activation.

Jurkat cells are an immortalized T-cell line derived from a patient with acute T-cell leukemia. They retain many characteristics of human T lymphocytes, including expression of the T-cell receptor (TCR)/CD3 complex, serving as a standard model for TCR signaling, apoptosis, and cytokine production. Their robust growth in suspension culture and ease of genetic manipulation make them ideal for generating knockout cell lines. In the BTN3A3 knockout context, the Jurkat background offers a well-defined platform to examine how butyrophilin proteins modulate T-cell responses independently of the complex interactions found in primary T-cell subsets.

BTN3A3 is a butyrophilin subfamily 3 protein that functions as a key phosphoantigen sensor, recognizing isopentenyl pyrophosphate (IPP). Upon phosphoantigen binding, BTN3A3 interacts with BTN2A1 to trigger conformational changes that activate V??9V??2 T cells through the TCR. Additionally, BTN3A3 provides co-stimulatory signals to ???? T cells, engaging downstream cascades including PI3K/AKT and MAPK pathways. These signals converge on NF-??B activation and promote IL-2 production. BTN3A3 is upregulated by cytokines such as IFN-?? and TNF-??, linking it to innate and adaptive immunity. In this knockout model, disruption of BTN3A3 is expected to impair phosphoantigen-driven signaling, reduce phosphorylation of ERK and AKT, and diminish cytokine secretion upon stimulation.

In Jurkat cells, which are widely used to dissect TCR-proximal signaling, BTN3A3 knockout creates a loss-of-function system to evaluate its contribution to T-cell activation thresholds. Since Jurkat cells lack ?æ? TCR, this model specifically highlights the co-signaling functions of BTN3A3 that intersect with ???? TCR pathways. Researchers can compare wild-type and BTN3A3-deficient Jurkat cells to assess changes in activation markers (e.g., CD69, CD25), phosphorylation kinetics of ERK and AKT, and IL-2 secretion. This defined system circumvents primary cell heterogeneity and enables high-throughput screening of modulators affecting BTN3A3-dependent signaling, including checkpoint inhibitors or phosphoantigen analogs.

This line is ideal for dissecting phosphoantigen recognition and T-cell co-regulation in autoimmune disease and cancer research. Key applications include investigating BTN3A3-dependent activation and tolerance, studying ?æ? T cell biology through co-culture with V??9V??2 cells, screening butyrophilin-targeted immune modulators, and validating BTN3A3 as an immunotherapy target. Standard assays include Western blotting and RT-qPCR for confirmation, flow cytometry for phospho-ERK/AKT and CD69/CD25, IL-2 ELISA, and proliferation. For further technical details, please contact Ascent Research.