The ALOX12B Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HeLa human cervical carcinoma cell line, providing a loss-of-function model for the ALOX12B gene. This product delivers a heterogeneous pool of edited cells with targeted disruption of ALOX12B, enabling robust functional genomics studies without the need for clonal isolation.
HeLa cells are an immortalized human epithelial line derived from a cervical adenocarcinoma, harboring integrated human papillomavirus type 18 (HPV18) sequences. They are one of the most extensively used cell models in biomedical research, particularly for cancer biology, signal transduction, and drug response studies. Their rapid proliferation, ease of culture, and well-characterized genomic landscape make them a versatile host for gene knockout experiments.
ALOX12B encodes arachidonate 12-lipoxygenase, 12R type, a key enzyme in the epidermal lipid barrier pathway. It catalyzes the oxygenation of arachidonic acid to 12R-hydroperoxyeicosatetraenoic acid (12R-HPETE), which is subsequently converted by ALOXE3 to bioactive hepoxilins, such as hepoxilin A3. ALOX12B activity is regulated by calcium influx, cytokines including IL-4 and IL-13, and nuclear receptors like PPAR and RAR/RXR, and it functionally interacts with fatty acid binding proteins and membrane phospholipids. Downstream, hepoxilins influence ceramide synthesis and the expression of keratinocyte differentiation markers, including involucrin and loricrin, thus maintaining skin barrier integrity.
In HeLa cells, which are of epithelial origin but not typically associated with epidermal differentiation, knockout of ALOX12B offers a distinct system to investigate eicosanoid signaling outside the skin context. Disruption of this gene may alter lipid mediator profiles, modulate ferroptosis sensitivity, and impact pathways relevant to cancer cell lipid metabolism. This model enables researchers to explore the ectopic functions of a skin-barrier enzyme in a tumorigenic background, providing insights into the intersection of lipid biology and oncogenic signaling.
These polyclonal knockout cells are applicable in a range of experimental settings, from mechanistic dissection of arachidonic acid metabolism to phenotypic screening for modulators of ferroptosis and ichthyosis-related pathways. Typical assays include RT-qPCR and western blotting for gene and protein expression analysis, LC?MS/MS lipidomics to profile eicosanoid changes, immunofluorescence for cellular localization studies, and cell viability assays to assess ferroptotic responses. The polyclonal nature allows study of diverse editing events, enhancing the robustness of population-level observations. For further information or technical inquiries, please contact Ascent Research.
