INSR Knockout HAP1 Cell Line

The choice depends on whether you are studying INSR (insulin receptor)'s role as the principal insulin-binding receptor tyrosine kinase or modeling type A and type B insulin resistance syndromes. The Knockout line is the standard tool for asking whether INSR is required for these processes — INSR is a receptor tyrosine kinase that, upon insulin binding, autophosphorylates and phosphorylates IRS1/IRS2 substrates, initiating PI3K-AKT signaling that drives glucose uptake, protein synthesis, and lipid storage. Overexpression is useful for studying INSR gain-of-function effects. Important consideration: IGF1R paralog expression analysis is essential — INSR and IGF1R can form heterodimers and share substantial substrate scope. This product complements the parallel IRS1 Knockout in HAP1 (also available) for upstream-downstream insulin signaling pathway dissection. Rescue with wild-type or kinase-dead INSR is the standard specificity control. The knockout is valuable for studying insulin resistance mechanisms, INSR-related diabetes syndromes (e.g., leprechaunism, Rabson-Mendenhall syndrome, type A insulin resistance), and emerging INSR-targeted drug development.

Primary applications: • Insulin binding and signaling: insulin-induced phospho-INSR (Y1162/Y1163), phospho-IRS1, and phospho-AKT analysis in INSR-null cells. • Severe insulin resistance modeling: rescue with patient-derived INSR mutations (e.g., leprechaunism, Rabson-Mendenhall syndrome variants) for genotype-function studies. • Insulin/IGF-1 receptor specificity: parallel IGF1R analysis to dissect INSR-specific versus IGF1R-shared functions. • Combined INSR+IRS1 dissection: parallel analysis with IRS1 Knockout in HAP1 (also available) for receptor-substrate dissection. EDITGENE recommends this model for researchers investigating insulin receptor biology, severe insulin resistance syndromes, and insulin-IGF signaling specificity.

Yes. INSR rescue experiments require attention to RTK architecture: • Construct design: use a codon-modified INSR sequence with a small intracellular C-terminal tag (FLAG, HA). INSR is processed from a single precursor into α and β chains (α2β2 tetramer) — preserve processing and membrane topology. • Surface localization validation: confirm plasma membrane α2β2 tetramer expression before functional assays. • Kinase-dead rescue: K1030A or K1018A in the ATP-binding lysine abolishes catalytic activity. • Insulin resistance syndrome mutation rescue: patient-derived INSR mutations enable disease genotype-function studies. • Functional readout: rescue should restore insulin-induced phospho-INSR, IRS1 phosphorylation, and downstream PI3K-AKT signaling. HAP1-specific considerations: • Diploidization: HAP1 cells gradually diploidize during extended culture — confirm ploidy by flow cytometry at the time of phenotypic assay. • Integration site sensitivity: position effects on transgene expression are more pronounced in near-haploid backgrounds; generating multiple independent rescue clones is strongly recommended. • Transduction efficiency: HAP1 transduces with lentivirus at moderate efficiency — increase MOI compared to standard immortalized lines.