PIK3R1 Knockout HAP1 Cell Line

The choice depends on whether you are studying PIK3R1 (p85α)'s role as the principal class IA PI3K regulatory subunit or modeling SHORT syndrome and immunodeficiency 36. The Knockout line is the standard tool for asking whether p85α is required for class IA PI3K function — p85α stabilizes p110 catalytic subunits, recruits PI3K to phosphotyrosine motifs through SH2 domains, and maintains p110 in an autoinhibited state until receptor activation. Overexpression is useful for studying p85α in heterologous contexts or for testing disease-associated mutations. For PI3K pathway research, the EDITGENE PIK3R1 Knockout in HAP1 is a workhorse mechanistic platform. PIK3R1 mutations cause multiple human diseases: SHORT syndrome (loss-of-function, R649W), agammaglobulinemia 7 (loss-of-function), and immunodeficiency 36 (activated PI3Kδ syndrome 2, gain-of-function from p85α-mediated p110δ disinhibition). Disease variant rescue enables genotype-function studies. This product complements the parallel PIK3R2 Knockout (also available).

Primary applications: • PI3K activity: phospho-AKT and downstream mTORC1 substrate analysis in the p85α-null background. • Multi-disease modeling: rescue with SHORT syndrome (R649W), agammaglobulinemia, or activated PI3Kδ syndrome 2-associated PIK3R1 mutations for genotype-function studies. • RTK-PI3K signaling: insulin, EGF, or PDGF-stimulated PI3K activation following p85α loss. • p110-binding studies: co-immunoprecipitation analysis of p110α/β/δ stability and binding in the absence of p85α. EDITGENE recommends this model for researchers investigating PI3K regulatory subunit biology, SHORT syndrome, agammaglobulinemia, and activated PI3Kδ syndrome.

Yes. p85α rescue experiments are well-established for PI3K research: • Construct design: use a codon-modified PIK3R1 sequence with a small C-terminal tag (FLAG, HA). p85α has SH3, two SH2 domains, BCR-homology domain, and iSH2 — preserve all elements. • Multi-disease mutation rescue: SHORT syndrome (R649W), agammaglobulinemia 7, and APDS2 (activated PI3Kδ syndrome 2) mutations enable comprehensive disease studies. • p110-binding-deficient rescue: iSH2 mutations enable distinguishing p110-stabilization from other p85α functions. • Functional readout: rescue should restore p110 stabilization, RTK recruitment, and class IA PI3K activity. 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.