PIK3R2 Knockout HAP1 Cell Line

The choice depends on whether you are studying PIK3R2 (p85β)'s role as a regulatory subunit of class IA PI3K or modeling MPPH (megalencephaly-polymicrogyria-polydactyly-hydrocephalus) syndrome. The Knockout line is the standard tool for asking whether p85β is required for class IA PI3K function — p85β complexes with p110α/β/δ catalytic subunits to form PI3K heterodimers, with p85α (PIK3R1) and p85β (PIK3R2) being the two principal regulatory isoforms. Overexpression is useful for studying p85β in heterologous expression contexts or for testing disease-associated mutations. Important consideration: p85α and p85β share substantial functional overlap — single PIK3R2 knockout may show modest phenotypes if p85α compensates. PIK3R2 mutations cause MPPH syndrome (megalencephaly-polymicrogyria-polydactyly-hydrocephalus) through gain-of-function effects on PI3K activity — disease variant rescue enables genotype-function studies. This product complements the parallel PIK3R1 Knockout in HAP1 (also available) for comprehensive PI3K regulatory subunit dissection.

Primary applications: • PI3K activity: phospho-AKT (S473, T308) Western blot to assess class IA PI3K activity in the absence of p85β. • MPPH disease modeling: rescue with patient-derived gain-of-function PIK3R2 mutations for genotype-function studies of megalencephaly-polymicrogyria-polydactyly-hydrocephalus syndrome. • Paralog studies: parallel comparison with PIK3R1 (p85α) Knockout in HAP1 (also available) for comprehensive PI3K regulatory subunit dissection. • PI3K inhibitor specificity: assessment of PI3K inhibitor sensitivity given altered p85α/p85β balance. EDITGENE recommends this model for researchers investigating PI3K regulatory subunit biology and MPPH syndrome mechanisms.

Yes. p85β rescue experiments require attention to PI3K regulatory subunit architecture: • Construct design: use a codon-modified PIK3R2 sequence with a small C-terminal tag (FLAG, HA). p85β has SH3 domain, two SH2 domains (N-SH2, C-SH2), and inter-SH2 (iSH2) domain that binds p110 — preserve all elements. • p110-binding-deficient rescue: iSH2 mutations disrupt p110 binding and serve as the standard control for p110-stabilization functions. • MPPH mutation rescue: gain-of-function PIK3R2 mutations enable disease genotype-function studies. • Functional readout: rescue should restore p110 stabilization and class IA PI3K activity following RTK stimulation. 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.