The HAVCR1 Knockout HCT 116 Polyclonal Cells consist of a polyclonal population of HCT 116 human colorectal carcinoma cells engineered via CRISPR/Cas9 to disrupt the HAVCR1 gene, thereby abolishing functional TIM?1 protein expression. This heterogeneous knockout pool preserves genetic diversity, offering a loss-of-function model free of clonal artifacts for investigating TIM?1-dependent processes in cancer and immune contexts.
The parental HCT 116 line is an epithelial cell model derived from a male colorectal carcinoma, featuring oncogenic KRAS G13D and CTNNB1 mutations and mismatch repair deficiency due to MSH2 mutation. Widely employed in colorectal cancer research, these cells are tumorigenic and serve as a standard system for drug screening, Wnt/???catenin pathway studies, and oncogenic signaling analyses.
HAVCR1 encodes T-cell immunoglobulin and mucin domain?1 (TIM?1), a phosphatidylserine (PtdSer) receptor and co?stimulatory molecule. TIM?1 is activated by PtdSer binding, TIM?4 engagement, or hepatitis A virus capsid proteins, and is regulated by upstream signals such as TCR/CD3 stimulation, IL?4/STAT6, NF???B, and TGF???1. Upon activation, TIM?1 recruits Src family kinases and propagates signaling via PI3K/AKT, MAPK, and NF???B cascades, also coupling to the TCR/CD3?CZAP70?CLAT?CPLC??1 axis. Downstream, it promotes Th2 cytokine expression (IL?4, IL?13) and modulates autophagy, phagocytosis, and cell survival. Thus, TIM?1 participates in T?cell co?stimulation, viral entry, efferocytosis, and kidney injury repair pathways.
In HCT 116 cells, HAVCR1 knockout disrupts TIM?1?mediated PtdSer recognition and downstream PI3K/AKT and NF???B signaling, potentially affecting apoptosis, autophagy, and immune crosstalk. This model allows dissection of TIM?1 functions within a defined oncogenic background, exploring intersections between PtdSer signaling, mutant KRAS, and ???catenin pathways in mismatch repair?deficient colorectal cancer.
Applications span virology (hepatitis A virus entry studies), immunology (T?cell co?stimulation and Th2 regulation), and cancer biology (drug sensitivity screening, migration/invasion assays, tumor microenvironment interactions). Standard assays include flow cytometry, western blotting, RT?qPCR, apoptosis assays, NF???B reporter assays, co?immunoprecipitation of interaction partners, and ELISA?based cytokine profiling. For inquiries, please contact Ascent Research.
