The HAVCR2 Knockout HEK293T Polyclonal Cells represent a ready-to-use CRISPR/Cas9-edited polyclonal knockout cell population, offering a heterogeneous loss-of-function model for the human HAVCR2 gene. This polyclonal pool provides a robust tool for studying TIM-3-mediated signaling without clonal selection, enabling researchers to interrogate immune checkpoint biology in a genetically tractable system. By disrupting HAVCR2, the product facilitates investigation of loss-of-function phenotypes across a mixed cellular background, supporting assays that require a diverse genotypic representation.
HEK293T cells are a widely utilized human embryonic kidney epithelial cell line immortalized by stable expression of the SV40 large T antigen. Their high transfectability, rapid proliferation, and capacity for efficient protein production make them an ideal host for gain- and loss-of-function studies. This background is particularly suited for experiments requiring exogenous expression of wild-type or mutant TIM-3 constructs, as well as for viral packaging and large-scale biochemical analyses, providing a reliable and well-characterized platform for dissecting gene function.
HAVCR2 encodes TIM-3, an immune checkpoint receptor expressed on exhausted T cells that plays a pivotal role in immune tolerance and T-cell dysfunction. Upon binding to ligands such as galectin-9 (LGALS9) or phosphatidylserine, TIM-3 recruits Src family kinases FYN and LCK, leading to the disruption of its interaction with the adaptor BAT3. This event inhibits proximal T-cell receptor (TCR) signaling, suppresses the transcriptional activity of NFATc1 and AP-1, and promotes apoptosis via caspase-3 activation. TIM-3 signaling is regulated by upstream factors including IL-27, T-bet, NFATc1, and STAT4, and it intersects with other immune regulators such as CEACAM1 and HMGB1 to modulate immune synapse stability and signal termination.
In the HEK293T background, knockout of HAVCR2 removes endogenous TIM-3 expression, creating a clean cellular environment to study the receptor??s signaling network through reconstitution experiments. Although HEK293T cells lack the full T-cell machinery, they support the mechanistic dissection of TIM-3 interactions and downstream pathways when co-expressed with relevant signaling components. This model is particularly valuable for analyzing ligand-induced phosphorylation events, protein complex formation, and transcriptional outputs without the confounding effects of endogenous TIM-3, enabling precise mapping of functional domains and signaling motifs.
This polyclonal knockout cell population is ideally suited for a range of research applications, including but not limited to investigating TIM-3 ligand-receptor interactions by co-immunoprecipitation and flow cytometry, assessing signaling pathway activity through NFAT/AP-1 reporter assays, and studying apoptotic responses via caspase-3 activation assays. It also supports Western blotting for protein expression analysis, immunocytochemistry for subcellular localization studies, and serves as a control for CRISPR validation experiments. By combining the versatility of HEK293T cells with targeted HAVCR2 disruption, this product facilitates comprehensive analyses of immune checkpoint mechanisms. For further information, please contact Ascent Research.
