STK38 Knockout HAP1 Cell Line

The choice depends on whether you are studying STK38's role in Hippo pathway-related signaling, centrosome duplication regulation, or apoptosis modulation. The Knockout line is the standard tool for asking whether STK38 (NDR1) is required for these processes — STK38 has been implicated in MOB1-mediated activation downstream of Hippo signaling. Overexpression is useful for testing sufficiency for substrate phosphorylation or for studying STK38's contribution to cancer biology contexts. For STK38 research, the EDITGENE Knockout in HAP1 is the more rigorous tool, though STK38L paralog compensation should be considered. Combined analysis with the parallel STK38L knockout (also available from EDITGENE) enables NDR family-specific dissection. Rescue with wild-type or kinase-dead STK38 is essential, particularly for studies of NDR kinase inhibitor specificity.

Primary applications: • Hippo pathway-related signaling: YAP/TAZ phosphorylation and substrate analysis downstream of NDR1 activation. • Apoptosis and stress response: cell death assays following pro-apoptotic stimuli to characterize NDR1's role in apoptotic signaling. • MOB1 binding studies: co-immunoprecipitation analysis of STK38-MOB1 complex formation, the key NDR activation step. • Paralog studies: combined analysis with STK38L knockout (also available) dissects family-specific functions. EDITGENE recommends this model for researchers investigating NDR1 kinase biology, Hippo signaling, and apoptotic regulation.

Yes. STK38 rescue experiments require attention to NDR family activation: • Construct design: use a codon-modified STK38 sequence with a small C-terminal tag (FLAG, HA). The N-terminal regulatory region and kinase domain are both essential. • Kinase-dead rescue: an active site mutation (K117A or D200A) serves as the catalytic activity specificity control. • MOB1-binding-deficient rescue: NTR mutations that disrupt MOB1 binding distinguish activated kinase functions from basal kinase activity. • Inhibitor specificity studies: NDR family inhibitors often show cross-reactivity — rescue with STK38 wild-type versus kinase-dead enables on-target validation. 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.