The HAVCR1 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population generated by disruption of the HAVCR1 gene in the HAP1 host cell background. This polyclonal knockout pool is designed for loss-of-function studies of the hepatitis A virus cellular receptor 1 (HAVCR1/TIM-1) without selection for specific clonal derivatives. The population-wide gene disruption provides a versatile tool for investigating TIM-1 biology in a near-haploid genetic environment.
HAP1 cells, derived from the KBM-7 male chronic myeloid leukemia line, are characterized by a near-haploid karyotype that simplifies genetic manipulation and facilitates efficient CRISPR/Cas9-mediated knockout generation. Their adherent growth and stable haploid state make them particularly suited for functional genomics, enabling researchers to analyze gene function in the absence of a second allele that might mask phenotypic effects. The chronic myeloid leukemia origin also provides a context for studying signaling pathways relevant to hematological malignancies.
HAVCR1 encodes TIM-1, a type I transmembrane glycoprotein serving as a receptor for phosphatidylserine and the hepatitis A virus capsid. In T cells, ligand engagement leads to LCK-mediated phosphorylation of the cytoplasmic tail, recruiting and activating PI3K?CAKT1 signaling. Concurrently, the MAPK/ERK cascade is stimulated via ZAP70 and PLCG1, resulting in NFATC1 activation and enhanced secretion of IL-2 and IL-4. Interacting partners include TIM-4 and phosphatidylserine; TIM-1 expression is upregulated by IL-4, TCR engagement, oxidative stress, and tissue injury. In kidney injury, the homolog KIM-1 mediates phagocytic uptake of apoptotic cells, a role conserved in human TIM-1.
Although HAP1 cells lack a full T-cell receptor signaling machinery, they express core components such as LCK and PI3K?CAKT pathway members, allowing reconstitution studies of TIM-1-mediated signaling events. The haploid background ensures that disruption of the single HAVCR1 allele results in a complete loss-of-function model without confounding compensation from a second allele. This clean genetic system is particularly advantageous for studying the intrinsic signaling properties of TIM-1, including lipid kinase activation and transcriptional regulation, without the complexity of T-cell-specific factors. The polyclonal nature of the population maintains experimental robustness by averaging clone-to-clone variation while preserving the knockout genotype at the pool level.
These polyclonal knockout cells provide a versatile platform for studying hepatitis A virus entry mechanisms, TIM-1-dependent phagocytic clearance, and regulatory T-cell signaling pathways. Standard assays including flow cytometry, western blotting, viral entry assays, and apoptotic cell uptake can be employed to assess TIM-1 function and downstream signaling. Additionally, the model is suitable for high-throughput screening of novel ligands or inhibitors targeting the TIM-1?Cphosphatidylserine interaction, and for investigating the role of TIM-1 in acute kidney injury models. For further technical inquiries, please contact Ascent Research.
