The HAVCR1 Knockout KYSE-30 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population with disrupted HAVCR1 gene expression, abolishing TIM-1 protein. This loss-of-function model enables functional interrogation of HAVCR1-dependent processes in a human esophageal cancer background without clonal selection artifacts, offering a robust system for signaling and genomic studies.
KYSE-30 is an adherent epithelial cell line derived from a well-differentiated esophageal squamous cell carcinoma in a 64-year-old male. These cells retain hyperactivated PI3K/AKT, MAPK/ERK, and NF-??B pathways characteristic of ESCC, making them an ideal host for HAVCR1 knockout studies. The polyclonal pool is generated by introducing CRISPR/Cas9 components and selection, resulting in a heterogeneous population that mirrors tumor genetic diversity and permits analysis of varying gene disruption levels.
HAVCR1 (TIM-1) is a transmembrane receptor for phosphatidylserine and hepatitis A virus that activates PI3K/AKT and MAPK/ERK signaling. Upstream regulation involves IL-4/IL-13-induced STAT6 and TCR-stimulated NFAT. Upon engagement, HAVCR1 promotes AKT1 and MTOR phosphorylation, upregulating pro-survival factors BCL2 and BIRC5 and cell cycle regulators CCND1 and EIF4EBP1. Concurrently, it stimulates MAPK1/ERK2 and NFKB1 through IKBKB and RELA. HAVCR1 interacts with TIMD4, LCK, ZAP70, and ITGB1, integrating immune and adhesion receptor signals to coordinate proliferation, survival, and immune modulation.
In KYSE-30 cells, HAVCR1 signaling sustains tumor cell proliferation, survival, and migration. CRISPR/Cas9-mediated HAVCR1 knockout disrupts AKT1 and ERK2 phosphorylation, attenuates NF-??B activation, and reduces expression of downstream targets such as CCND1 and BCL2L1, potentially impairing viability and invasiveness. The polyclonal knockout population models heterogeneous gene loss, avoiding clonal artifacts and enabling investigation of dosage-dependent phenotypic effects.
Research applications include signal transduction analysis via phospho-western blotting (p-AKT, p-ERK) and kinase arrays, proliferation and migration assays (MTT, transwell), apoptosis detection (annexin V flow cytometry), and protein interaction studies (co-immunoprecipitation, immunofluorescence with TIMD4 or LCK). This model supports esophageal squamous cell carcinoma biology, immune evasion studies, drug target validation, and viral entry research. For further technical details or experimental design assistance, please contact Ascent Research.
