The HAVCR2 Knockout HeLa Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal cell population generated from the human HeLa cervical adenocarcinoma line, with targeted disruption of the HAVCR2 gene (encoding TIM-3). This loss-of-function model is designed for studying immune checkpoint-independent roles of TIM-3 in epithelial cancer cell signaling, apoptosis, and survival.
HeLa cells are an HPV18-positive, epithelial-like immortalized line widely used in cancer biology. Their robust growth and well-characterized signaling networks make them a standard platform for examining gene function via CRISPR-based knockout, providing a relevant context for cervical cancer research.
HAVCR2/TIM-3 is an immune checkpoint receptor that interacts with ligands such as galectin-9, HMGB1, and CEACAM1. In non-immune cells, it modulates PI3K/Akt, NF-??B, and MAPK/ERK pathways, with downstream effects on cell cycle regulators (p21, Cyclin D1) and apoptosis mediators (Bax, Bak). Upstream regulators include cytokines (IL-2, IL-15, TGF-??) and transcription factors (NFAT, STAT3, NF-??B). TIM-3 also associates with adaptor proteins PI3K p85, Grb2, and Src kinase Fyn.
Knockout of HAVCR2 in HeLa cells allows dissection of TIM-3 function outside the immune system, particularly its impact on apoptotic susceptibility and proliferative signaling. The polyclonal nature of the population captures diverse editing outcomes, avoiding clonal selection bias and better representing heterogeneous tumor cell responses.
Applications include validating TIM-3 as a cancer target, studying galectin-9/TIM-3-mediated signaling, and screening compounds that modulate these pathways. Typical assays include Western blot for phospho-Akt/ERK/NF-??B, Annexin V apoptosis and MTT proliferation analysis, Transwell migration, and co-culture with immune cells. For further details, contact Ascent Research.
