KLRB1 Knockout NCI-H1299 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of NCI-H1299 human non-small cell lung carcinoma epithelial cells carrying a targeted disruption of the KLRB1 gene. This gene encodes CD161, a C-type lectin receptor that regulates natural killer and T cell effector functions. The polyclonal format provides a heterogeneous pool of edited cells, enabling robust loss-of-function studies without clonal variability. The product is supplied as an adherent epithelial cell population, ready for downstream functional and molecular analyses.
The NCI-H1299 cell line, derived from a lymph node metastasis of lung adenocarcinoma, serves as a standard model of non-small cell lung carcinoma. These adherent epithelial cells are p53-null due to a homozygous partial deletion of TP53, facilitating studies of genomic instability and apoptosis resistance. Widely used in cancer biology and drug screening, NCI-H1299 cells retain key adenocarcinoma characteristics and are amenable to gene editing, making them a robust host for knockout studies.
KLRB1 encodes CD161, a C-type lectin receptor that binds LLT1 (CLEC2D) and recruits SHP-1 and SHP-2 phosphatases to deliver inhibitory signals. This axis suppresses NK and T cell cytotoxicity and pro-inflammatory cytokine secretion, including IFN-gamma and TNF-alpha. KLRB1 expression is induced by IL-2, IL-12, and IL-15 and is transcriptionally regulated by T-bet and Eomes. The SHP-1/SHP-2-mediated dephosphorylation of downstream targets attenuates activating receptor signaling, establishing CD161 as an immune checkpoint that dampens immune responses. Disruption of KLRB1 potentially unleashes effector functions, augmenting anti-tumor immunity.
In NCI-H1299 lung carcinoma cells, KLRB1 knockout provides a model to dissect the CD161-LLT1 axis within the tumor microenvironment. Although CD161 is primarily on immune cells, its tumor expression can impact interactions with LLT1-bearing stromal cells. KLRB1 disruption in this p53-null line allows investigation of altered signaling, cytokine output, and immune-mediated killing. Co-cultures with NK or T cells can assess whether KLRB1 ablation in tumor cells improves recognition and lysis, mirroring checkpoint blockade. This model aids in evaluating CD161’s role in immune evasion in non-small cell lung cancer.
Typical applications involve western blotting and RT-qPCR to verify KLRB1 disruption, flow cytometric analysis of surface CD161, and functional co-culture killing assays using NK cells or T cells to measure tumor cell lysis. Cytokine secretion profiling by ELISA, especially for IFN-gamma and TNF-alpha, can reveal shifts in immune effector output. The polyclonal knockout population is also well suited for screening compounds that target the CD161-LLT1 interaction and for dissecting downstream signaling pathways. For additional information or to request a quote, please contact Ascent Research.
