PPP2R5E Knockout HAP1 Cell Line

The choice depends on whether you are studying PPP2R5E (B56ε, PR61ε)'s role as a B' regulatory subunit of PP2A determining substrate specificity or its specific contributions distinct from other B56 paralogs. The Knockout line is appropriate for asking whether B56ε is required for PP2A targeting to specific substrates — B56ε is one of five B56 isoforms (α, β, γ, δ, ε encoded by PPP2R5A-E) that determine PP2A holoenzyme substrate selectivity. Overexpression is useful for studying B56ε in heterologous expression contexts. Important consideration: B56α-ε paralogs have substantial substrate overlap — single PPP2R5E knockout in HAP1 may show modest phenotypes if other B56 isoforms compensate. Rescue with wild-type PPP2R5E is the standard specificity control. This product complements the parallel B56α, B56B, and B56D knockouts (also available) for comprehensive B56 family functional dissection.

Primary applications: • PP2A holoenzyme assembly: co-immunoprecipitation analysis of PPP2CA (catalytic C) and PPP2R1A (scaffold A) with B56ε to characterize PP2A-B56ε holoenzyme composition. • B56ε-specific substrate identification: phosphoproteomics in the knockout to identify substrates preferentially dephosphorylated by PP2A-B56ε. • B56 family comparative studies: parallel analysis with other PPP2R5 family knockouts for B56 paralog-specific function characterization. • PP2A reactivator response: NZ-8-061 (SMAP) and other PP2A reactivator specificity testing. EDITGENE recommends this model for researchers investigating PP2A B' regulatory subunit specialization and B56ε-specific substrate biology.

Yes. PPP2R5E rescue experiments require attention to PP2A holoenzyme assembly: • Construct design: use a codon-modified PPP2R5E sequence with a small C-terminal tag (FLAG, HA). B56ε contains conserved B56 domain — preserve scaffold subunit (Aα/PPP2R1A) binding region. • Holoenzyme assembly validation: co-immunoprecipitation with PPP2CA and PPP2R1A confirms B56ε-PP2A holoenzyme formation. • Substrate-binding-deficient rescue: mutations in the conserved B56 substrate-recruitment surface enable distinguishing scaffolding from substrate targeting functions. • Functional readout: rescue should restore B56ε-PP2A holoenzyme assembly and B56ε-dependent substrate dephosphorylation. 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.