The ALOX15 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line, with targeted disruption of the ALOX15 gene. This polyclonal model provides a loss-of-function system for studying arachidonate 15-lipoxygenase, a key enzyme in lipid mediator biosynthesis. The heterogeneous editing in the population enables robust interrogation of ALOX15-dependent phenotypes without clonal selection.
HCT 116 is a well-established colorectal carcinoma epithelial cell line featuring microsatellite instability (MSI) and oncogenic mutations in KRAS (G13D) and PIK3CA (H1047R). These alterations activate MAPK and PI3K/AKT signaling, driving tumorigenic properties such as proliferation, survival, and migration. The cell line serves as a standard model for colorectal cancer research, including studies on drug sensitivity and tumor microenvironment interactions.
ALOX15 encodes arachidonate 15-lipoxygenase, which oxygenates arachidonic acid to 15-HPETE; this is reduced by GPX4 to 15-HETE, a signaling lipid that influences inflammation, apoptosis, and ferroptosis. ALOX15 expression is regulated by IL-4/IL-13?CSTAT6 signaling, PPAR??, NF-??B, and p53. Its activity interacts with COX-2, ALOX5, and phospholipase A2, and its product 15-HETE can modulate PPAR??, MAPK, and PI3K/AKT pathways. Additionally, ALOX15 contributes to lipoxin A4 synthesis, linking it to resolution of inflammation and caspase-mediated apoptosis.
In colorectal cancer, ALOX15 impacts lipid peroxidation dynamics and ferroptosis susceptibility, which are critical for tumor cell fate. The HCT 116 background, with its activated oncogenic pathways, provides an ideal context to assess how loss of ALOX15 alters ferroptosis sensitivity to agents like RSL3 and erastin, and how it reshapes the lipid mediator landscape. This knockout model facilitates the exploration of ALOX15’s role in colorectal tumor progression and therapeutic response.
These polyclonal knockout cells are suited for assays such as C11-BODIPY lipid peroxidation measurements, ferroptosis induction with RSL3/erastin, cell viability testing, Western blotting for ALOX15, RT-qPCR, migration and invasion assays, colony formation, and LC-MS/MS lipid mediator profiling. They enable dissection of arachidonic acid metabolism crosstalk with oncogenic signaling and support drug screening for ferroptosis modulators in colorectal cancer. For technical inquiries, please contact Ascent Research.
