The ALOX12 Knockout LoVo Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the ALOX12 gene. This product is a heterogeneous pool of LoVo cells harboring targeted gene disruption at the ALOX12 locus, generated without single-cell cloning, and provides a robust model to interrogate ALOX12-dependent signaling pathways. The polyclonal format enables functional analysis in a context that more closely reflects the genetic diversity of a tumor cell population, making it suitable for high-content screens, pooled functional genomics, and pathway dissection in colorectal cancer research.
Derived from the LoVo human colorectal adenocarcinoma cell line, this knockout model is established in a well-characterized host background originating from a metastatic lymph node of a Dukes’ type C colorectal adenocarcinoma patient. LoVo cells harbor oncogenic KRAS G13D and tumor-suppressive APC mutations, recapitulating key molecular features of aggressive colorectal cancer. The epithelial morphology and metastatic origin make these cells an invaluable system for studying invasion, proliferation, and signal transduction in a clinically relevant setting.
ALOX12 encodes arachidonate 12-lipoxygenase, which catalyzes the dioxygenation of arachidonic acid to produce 12-hydroperoxyeicosatetraenoic acid (12-HPETE) and its reduced metabolite 12-hydroxyeicosatetraenoic acid (12-HETE). These lipid mediators function as ligands for the G protein-coupled receptor GPR31, activating downstream ERK and protein kinase C (PKC) signaling, and contributing to NF-??B-mediated transcriptional responses. ALOX12 activity also suppresses glutathione peroxidase 4 (GPX4) and promotes ferroptosis by driving lipid peroxidation, in part through crosstalk with System xc- and COX-2. Upstream, ALOX12 expression is regulated by cytokines such as IL-4 and IL-13, as well as by KRAS, STAT3, and NF-??B, placing it at the intersection of inflammatory and oncogenic cascades.
In the LoVo cell model, ALOX12 knockout disrupts the generation of 12-HETE and its downstream effects on proliferation, migration, and ferroptosis sensitivity. Given the co-occurrence of KRAS and APC mutations, these polyclonal knockout cells provide a physiologically pertinent system to examine how ALOX12-mediated eicosanoid signaling intersects with hyperactivated MAPK/ERK and altered Wnt/??-catenin pathways. This model enables dissection of lipid-mediated regulatory mechanisms that may contribute to colorectal cancer progression and therapy resistance.
Typical research applications include ferroptosis mechanism studies, where ALOX12-dependent lipid peroxidation can be evaluated using C11-BODIPY probes and erastin/RSL3 induction; signal transduction analyses employing western blotting and RNA-seq to profile ERK, NF-??B, or GPX4 alterations; and migration assays to assess metastatic potential. Quantitative measurement of 12-HETE by ELISA, flow cytometric sorting of knockout populations, and RT-qPCR validation of target disruption are routinely employed. For further information, please contact Ascent Research.
