The ALOX12 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 786-O human clear cell renal cell carcinoma epithelial cell line. This gene-edited product features targeted disruption of the ALOX12 gene, enabling loss-of-function studies of the 12-lipoxygenase enzyme. The polyclonal format provides a heterogeneous population of edited cells, suitable for modeling genetic ablation without clonal selection artifacts.
The 786-O parental cell line was established from a primary clear cell adenocarcinoma of the kidney and is widely employed as a model system for renal cell carcinoma (RCC). 786-O cells exhibit characteristic epithelial morphology and harbor genetic alterations commonly found in RCC, including constitutive activation of HIF pathways. This background is ideal for investigating oncogenic signaling and tumor cell behavior in a clinically relevant context.
ALOX12 encodes an oxygenase that catalyzes the conversion of arachidonic acid to 12(S)-hydroperoxyeicosatetraenoic acid (12-HPETE), which is rapidly reduced to 12-hydroxyeicosatetraenoic acid (12-HETE). 12-HETE functions as a lipid mediator, binding to the G-protein-coupled receptor GPR31 to activate downstream pathways such as MAPK/ERK and PI3K/Akt. Transcriptional regulation of ALOX12 involves NF-??B and Sp1, while cytokines IL-4 and IL-13 can modulate its expression. Key interacting factors include phospholipase A2 (PLA2), which liberates arachidonic acid from membrane phospholipids, and calcium, which is required for ALOX12 activity. In the pathway, PLA2G4A supplies substrate, and PTGS2 represents a parallel branch of arachidonic acid metabolism.
In the 786-O RCC model, ALOX12-derived 12-HETE has been implicated in promoting tumor cell proliferation, migration, and invasive capacity through autocrine and paracrine mechanisms. By disrupting ALOX12, these polyclonal knockout cells allow researchers to dissect the contribution of the 12-HETE/GPR31 axis to malignant phenotypes and to assess the role of lipoxygenase-mediated signaling in the tumor microenvironment. This model is also relevant for understanding the crosstalk between inflammatory lipid mediators and oncogenic pathways.
Typical applications include comparative analyses of proliferation, migration, and invasion between wild-type and knockout populations, using assays such as MTT, Boyden chamber, and Annexin V-based apoptosis detection. Downstream signaling can be evaluated by phospho-ERK immunoblotting, while transcriptomic changes may be explored via RNA-seq. The model is further suited for pharmacological studies testing lipoxygenase inhibitors or assessing the impact of 12-HETE reconstitution. Quantification of 12-HETE by ELISA provides direct confirmation of disrupted enzymatic output. For further technical details, please contact Ascent Research.
