PLCG1 Knockout HAP1 Cell Line

The choice depends on whether you are studying PLCG1's role as the principal RTK-activated phospholipase C-γ isoform or its functions in T cell receptor signaling, growth factor responses, and cancer biology. The Knockout line is the standard tool for asking whether PLCγ1 is required for these processes — PLCγ1 is recruited to phosphotyrosine motifs through its SH2 domains, where receptor tyrosine kinases (EGFR, PDGFR, FGFR) and Src-family kinases activate it to hydrolyze PIP2 into IP3 and DAG. Overexpression is useful for studying PLCγ1 in cancer contexts where activating mutations occur (notably in cutaneous T-cell lymphoma and angiosarcoma). For RTK and TCR signaling research, the EDITGENE PLCG1 Knockout in HAP1 is a workhorse mechanistic platform — HAP1 supports clean loss-of-function studies of PLCγ1's contribution to RTK signaling. PLCG2 paralog expression should be assessed given functional overlap. Rescue with wild-type, lipase-dead (H335A), or cancer-associated activating mutant (R707Q, S345F, E1163K) PLCγ1 enables comprehensive structure-function studies. The knockout is valuable for studying PLCγ1-targeted compounds and PLCγ1-related cancer biology.

Primary applications: • RTK signaling: PIP2 hydrolysis, IP3 generation, and intracellular Ca²⁺ mobilization following EGF, PDGF, or FGF stimulation in PLCG1-null versus rescued cells. • DAG-PKC signaling: phospho-PKC substrate analysis given DAG's role as PKC activator. • Cancer biology: rescue with activating PLCG1 mutations (R707Q in cutaneous T-cell lymphoma, S345F, E1163K) for genotype-function studies of cancer-associated PLCγ1 activation. • PLC family comparison: PLCG2 paralog expression analysis for paralog-specific characterization. EDITGENE recommends this model for researchers investigating RTK-PLC signaling, cancer-associated PLCγ1 activation, and PLC-mediated calcium signaling.

Yes. PLCγ1 rescue experiments are well-established for receptor tyrosine kinase signaling research: • Construct design: use a codon-modified PLCG1 sequence with a small C-terminal tag (FLAG, HA). PLCγ1 has PH domain, EF hands, X-Y catalytic domains, SH2-SH2-SH3 array (specific to γ isoforms), and C2 domain — preserve all elements. • Lipase-dead rescue: the H335A mutation in the conserved catalytic histidine abolishes phospholipase activity and is the standard specificity control. • SH2 domain-mutant rescue: SH2 domain mutations disrupt phosphotyrosine binding and RTK recruitment without affecting intrinsic catalytic activity. • Cancer-associated activating mutation rescue: R707Q (CTCL hot spot) and other activating mutations enable disease genotype-function studies. • Functional readout: rescue should restore PIP2 hydrolysis (IP3 mass), Ca²⁺ mobilization, and downstream PKC activation 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.