The ALOX12 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population engineered for loss-of-function studies of the ALOX12 gene. This product is supplied as a heterogeneous pool of HAP1 cells harboring site-specific gene disruptions introduced by CRISPR/Cas9-mediated genome editing, providing a versatile model to interrogate ALOX12-dependent biology without clonal selection artifacts. The polyclonal format preserves population-level diversity while ensuring robust target-gene inactivation across the culture, making it suitable for bulk biochemical, functional, and pharmacological analyses. Researchers can employ this tool to dissect the roles of 12-lipoxygenase in signal transduction, lipid mediator production, and cellular stress responses.
The host cell line, HAP1, is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid karyotype??retaining only one copy of most chromosomes??greatly simplifies loss-of-function genetic manipulations and has made it a workhorse for genome-wide screens, drug-target validation, and pathway mapping. HAP1 cells exhibit a stable, adherent growth morphology and retain key signaling networks relevant to cancer biology and inflammation, offering a genetically tractable human background in which to study ALOX12 function without the confounding effects of redundant alleles present in diploid lines.
ALOX12 encodes arachidonate 12-lipoxygenase, a non-heme iron-containing dioxygenase that catalyzes the stereospecific oxygenation of arachidonic acid to 12-hydroperoxyeicosatetraenoic acid (12-HPETE), which is rapidly reduced by glutathione peroxidases to 12-hydroxyeicosatetraenoic acid (12-HETE). 12-HETE acts as a potent lipid mediator by engaging the G-protein-coupled receptor GPR31, triggering downstream cascades such as MAPK, PI3K/Akt, and NF-??B, and modulating cellular proliferation, migration, and oxidative stress responses. ALOX12 expression is regulated by cytokines (IL-4, IL-13), growth factors (EGF), and transcription factors (SP1, AP-1, NF-??B), and it functionally interacts with phospholipases A2, 5-lipoxygenase, and COX-2. Additionally, crosstalk with p53 and NADPH oxidases links ALOX12 to ferroptotic and redox-sensitive cell death pathways.
In the HAP1 background, disruption of ALOX12 enables dissection of its signaling contributions in a simplified genetic context, where phenotypic consequences can be observed without interference from a second functional allele. This model is particularly valuable for exploring the interplay between lipoxygenase-driven eicosanoid production and oncogenic signaling, as HAP1 retains central pathways relevant to leukemia and inflammation. By abolishing 12-HETE biosynthesis, the knockout cells allow direct assessment of ALOX12-dependent processes, including GPR31-mediated signaling, redox balance, and apoptosis regulation, providing a clean loss-of-function system for mechanistic and therapeutic investigations.
Key applications include profiling of lipid mediators by LC?CMS/MS, evaluating ferroptosis sensitivity under oxidative stress conditions, and performing co-culture assays to measure immune cell activation and platelet aggregation responses. The cells can be utilized in high-throughput screening formats for ALOX12 inhibitor discovery, Western blot and RT-qPCR validation of downstream targets, and inflammatory cytokine profiling. This tool supports research into cancer, cardiovascular disease, inflammatory disorders, and ferroptosis, offering a defined platform for deciphering the multifaceted roles of 12-lipoxygenase. For additional information or to discuss custom requirements, please contact Ascent Research.
