The KLRB1 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 786-O human renal cell line. This product provides a heterogeneous pool of cells with targeted disruption of the KLRB1 gene, enabling loss-of-function studies without clonal selection. The polyclonal format preserves genetic diversity while ensuring robust knockout representation, making it suitable for population-level analyses of KLRB1-dependent processes.
The parental 786-O cell line is a well-characterized model of clear cell renal cell carcinoma (ccRCC), originating from a primary adenocarcinoma. These cells harbor a mutant VHL gene, leading to constitutive hypoxia-inducible factor (HIF) activation, a hallmark of ccRCC tumorigenesis. Cultured as adherent epithelial cells, 786-O cells maintain key oncogenic signaling pathways and are widely used for renal cancer research, including studies on metabolic reprogramming and immune microenvironment interactions.
KLRB1 encodes CD161, an inhibitory C-type lectin receptor that modulates immune cell function. Upon engagement with its ligand CLEC2D (LLT1), KLRB1 undergoes tyrosine phosphorylation of its immunoreceptor tyrosine-based inhibitory motif (ITIM), recruiting the phosphatases SHP-1 and SHP-2. These phosphatases dephosphorylate proximal signaling molecules such as ZAP70 and the adaptor LAT, thereby dampening downstream activation cascades, including NFAT and NF-??B pathways. KLRB1 signaling is regulated by cytokines like IL-12, IL-18, and IL-23, as well as TCR/CD3 stimulation, positioning it as a critical checkpoint in NK and T cell effector responses.
In the context of 786-O cells, KLRB1 expression may contribute to immune evasion mechanisms. Renal tumors often upregulate ligands for inhibitory receptors, and the interaction between tumor-expressed CLEC2D and immune cell KLRB1 can suppress anti-tumor immunity. Engineered knockout of KLRB1 in this VHL-mutant renal carcinoma background allows researchers to dissect how loss of this receptor alters tumor-immune crosstalk, cytokine production, and downstream signaling in a cell-autonomous manner, or when co-cultured with immune effector cells.
These polyclonal knockout cells are ideal for functional assays such as co-culture experiments with NK cells or T cells to assess cytotoxicity and cytokine secretion. Researchers can employ flow cytometry to verify KLRB1 loss, phospho-flow to monitor SHP-1/SHP-2 recruitment, and Western blotting to evaluate signaling molecule phosphorylation. Additional applications include studying the impact of KLRB1 disruption on LLT1-mediated immune evasion, investigating NK cell-mediated cytotoxicity pathways, and screening for immune checkpoint modulators in renal cancer. For further technical details and ordering information, please contact Ascent Research.
