The ALOX12 Knockout A2780 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population designed for targeted disruption of the ALOX12 gene in the human A2780 ovarian carcinoma epithelial cell line. This polyclonal pool contains a heterogeneous mixture of edited alleles generated by Cas9-mediated DNA cleavage, providing a robust loss-of-function model without the clonal selection artifacts that can arise in monoclonal isolates. The product is supplied as a ready-to-use population that enables efficient screening of ALOX12-dependent phenotypes in a disease-relevant cellular context.
The host A2780 cell line was established from an untreated patient with ovarian endometrioid adenocarcinoma and serves as a standard in vitro model for epithelial ovarian cancer. These adherent cells maintain key hallmarks of ovarian carcinoma, including hormone responsiveness and characteristic chromosomal aberrations, making them particularly valuable for oncogenic signaling studies and drug sensitivity testing. Their well-documented growth properties and susceptibility to chemotherapeutic agents like cisplatin render them a reliable platform for dissecting mechanisms of intrinsic and acquired drug resistance.
ALOX12 encodes arachidonate 12-lipoxygenase, which catalyzes the dioxygenation of arachidonic acid to 12-hydroperoxyeicosatetraenoic acid (12-HPETE). This intermediate is rapidly reduced to 12-hydroxyeicosatetraenoic acid (12-HETE), a potent lipid mediator that engages the GPR31 receptor to activate downstream effectors including NADPH oxidase and NF-??B, while promoting integrin-mediated adhesion. ALOX12 expression is transcriptionally regulated by SP1 and is induced by IL-4, IL-13, and calcium influx through MAP kinase signaling. The enzyme operates in concert with cytosolic phospholipase A2, which mobilizes arachidonic acid from membrane phospholipids, and can crosstalk with 5-lipoxygenase within the eicosanoid network. In the ferroptosis pathway, ALOX12-generated lipid peroxides are integrated with GPX4, ACSL4, and LPCAT3 to influence cell death sensitivity.
In A2780 ovarian cancer cells, ALOX12 has been implicated in promoting proliferation and metastatic behavior through the 12-HETE/GPR31 signaling axis, which can stimulate NF-??B transcriptional programs and enhance cell motility. Additionally, ALOX12 activity modulates the cellular lipid peroxide landscape, potentially contributing to ferroptosis resistance??a critical determinant of tumor cell survival under oxidative stress. Disruption of ALOX12 in this background therefore allows direct interrogation of its role in ovarian tumor progression, metastasis, and therapeutic vulnerability, particularly in the context of standard-of-care platinum-based therapies.
These polyclonal knockout cells are tailored for a broad spectrum of applications, including mechanistic studies of eicosanoid signaling, ferroptosis induction assays with Erastin, proliferation and migration assessments via MTT or Transwell assays, and quantitative analysis of 12-HETE production by ELISA. They facilitate validation of ALOX12 as a therapeutic target and enable screening of compounds that modulate the lipoxygenase pathway. For further technical inquiries or custom solutions, researchers are encouraged to contact Ascent Research.
