The KLRB1 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the KLRB1 gene in the HCT 116 human colorectal carcinoma epithelial cell line. This polyclonal pool provides a loss-of-function model for studying CD161 (NKR-P1A) receptor biology without the limitations of clonal selection, enabling robust functional interrogation of KLRB1-dependent signaling in a genetically heterogeneous population. The knockout is achieved through targeted gene disruption, generating a versatile tool for immunological and oncological research applications.
The HCT 116 host cell line is a widely characterized human colorectal carcinoma epithelial model harboring an activating KRAS G13D mutation and a stabilizing ??-catenin (CTNNB1) mutation, which drive constitutive MAPK and Wnt pathway signaling. This genetic background recapitulates key features of aggressive colorectal tumors, including deregulated proliferation, survival, and invasive potential. The epithelial origin of HCT 116 cells renders them particularly suitable for examining tumor-intrinsic immune modulatory mechanisms and for co?culture studies with immune effector cells in a colorectal cancer context.
KLRB1 encodes the C-type lectin-like receptor CD161, which primarily interacts with its ligand CLEC2D (LLT1) to modulate immune cell activation. In NK cells, CD161 engagement can transmit inhibitory signals through ITIM-mediated recruitment of SHP-1 and SHP-2 phosphatases or activating signals via DAP12-coupled Syk and ZAP70 kinases, leading to PLC-?? activation and downstream PI3K-AKT, NF-??B, and MAPK pathway modulation. Upstream regulators such as IL-15, TGF-??, TBX21, and Eomes transcriptionally control KLRB1 expression, while downstream targets include IFNG and IL17A. The KLRB1?CCLEC2D axis influences cytotoxicity and cytokine production, making it a critical checkpoint in innate and adaptive immunity.
In the colorectal tumor microenvironment, ectopic or aberrant expression of CD161 on HCT 116 cells and surrounding immune infiltrates may promote immune evasion through inhibitory signaling or altered cytokine networks. Disruption of KLRB1 in this polyclonal HCT 116 model allows researchers to dissect the direct contribution of CD161 to tumor?Cimmune cell interactions, SHP-1/SHP-2-dependent signal transduction, and transcriptional programs that regulate inflammatory mediators. The absence of CD161 eliminates LLT1?mediated crosstalk, providing a clean background to evaluate tumor cell?Cintrinsic versus immune cell?Cextrinsic roles of this receptor?Cligand pair.
This knockout cell population is suitable for a broad range of applications, including immune evasion mechanism studies, tumor?CNK cell co?culture cytotoxicity assays, screening of immunomodulatory compounds, and functional analysis of CD161 in non?immune cells. Representative assays include flow cytometry for CD161 expression, RT?qPCR for KLRB1 mRNA, western blot for phospho?SHP?1, ELISA for IFN??? and IL?17, transwell migration/invasion assays, and immunofluorescence for LLT1 localization. For further technical specifications or batch?specific details, please contact Ascent Research.
