ALOX12 Knockout HAP1 Cell Line

The choice depends on whether you are studying ALOX12 (platelet-type 12-lipoxygenase, 12-LOX)'s role as the principal 12-HETE-generating enzyme or modeling its functions in platelet biology, ferroptosis, and emerging cancer applications. The Knockout line is the standard tool for asking whether 12-LOX is required for these processes — ALOX12 catalyzes the conversion of arachidonic acid to 12(S)-HPETE/12(S)-HETE; ALOX12 is highly expressed in platelets where 12-HETE contributes to platelet activation, and has emerging roles in ferroptosis (12-LOX-generated lipid peroxides drive iron-dependent cell death). Overexpression is useful for studying ALOX12 gain-of-function effects. Important consideration: ALOX12 (12S-LOX) is distinct from ALOX12B (12R-LOX, epidermal); these enzymes generate stereochemically opposite products from arachidonic acid. For platelet and ferroptosis research, the EDITGENE ALOX12 Knockout in HAP1 enables study of 12S-LOX biology. This product complements the parallel ALOX12B Knockout in HEK293 (also available, epidermal isoform) for stereochemistry dissection. Rescue with wild-type or catalytically-dead ALOX12 enables structure-function studies. The knockout is a critical specificity tool for ⭐ ML355 (selective ALOX12 inhibitor), and emerging ferroptosis-targeted and platelet-targeted therapeutics.

Primary applications: • Platelet activation: in heterologous platelet-relevant contexts, 12(S)-HETE production analysis given ALOX12's platelet enrichment. • Ferroptosis biology: lipid peroxidation analysis given 12-LOX's role in ferroptosis-driving lipid peroxides. • ALOX12 inhibitor specificity: critical genetic control for ⭐ ML355 (selective ALOX12 inhibitor) in emerging anti-platelet and ferroptosis-targeted drug development. • ALOX12B stereochemistry dissection: paired analysis with ALOX12B KO in HEK293 (also available) for 12S vs 12R-HETE stereochemistry studies. EDITGENE recommends this model for researchers investigating 12-lipoxygenase biology and emerging ferroptosis/anti-platelet therapeutic development.

Yes. ALOX12 rescue experiments are well-established for lipoxygenase research: • Construct design: use a codon-modified ALOX12 sequence with a small C-terminal tag (FLAG, HA). ALOX12 has the canonical lipoxygenase architecture with active site iron — preserve all elements. • Catalytically-dead rescue: iron-binding histidine cluster mutations abolish 12-LOX activity. • ML355-resistant rescue: gatekeeper mutations can confer ML355 resistance for on-target validation. • Functional readout: rescue should restore 12(S)-HETE generation from arachidonic acid. 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.