The SELENOI Knockout HCT 116 Cell Line is a CRISPR/Cas9-edited human colorectal carcinoma cell line featuring a targeted disruption of the SELENOI gene, encoding ethanolaminephosphotransferase. This loss-of-function model enables studies of SELENOI-dependent processes in a genetically defined epithelial cancer background. Derived from the widely used HCT 116 parental line, this knockout cell line provides a stable and renewable resource for investigating phosphatidylethanolamine synthesis and GPI anchor biology, suitable for a range of functional and phenotypic assays relevant to cancer metabolism and membrane dynamics.
The HCT 116 host cell line is a colorectal carcinoma epithelial model characterized by microsatellite instability (MSI), an activating KRAS G13D mutation, and constitutive ??-catenin signaling. These features recapitulate key oncogenic pathways active in a subset of colorectal cancers, making HCT 116 a standard system for dissecting tumor cell signaling, drug responses, and metabolic adaptations. Its adherent growth and epithelial morphology facilitate imaging, biochemical, and genetic manipulation. Combined with SELENOI knockout, this background allows targeted exploration of phospholipid metabolism in the context of colorectal tumorigenesis.
SELENOI catalyzes the final step in phosphatidylethanolamine (PE) synthesis via the CDP-ethanolamine branch of the Kennedy pathway, transferring phosphoethanolamine from CDP-ethanolamine to diacylglycerol. It functions downstream of ethanolamine kinase (ETNK1) and phosphoethanolamine cytidylyltransferase (PCYT2) and interacts with choline/ethanolamine phosphotransferase 1 (CEPT1). PE produced by SELENOI serves as a substrate for GPI anchor biosynthesis, acting upstream of PIGA and PIGK, and contributes to autophagosome membrane formation. SELENOI expression is regulated by transcription factors SREBF1 and PPARG in response to nutrient-sensitive signaling, linking membrane lipid homeostasis to cellular metabolic status.
In HCT 116 colorectal cancer cells, SELENOI disruption impairs PE synthesis and GPI anchor attachment, potentially altering surface expression of GPI-anchored proteins involved in cell adhesion, signaling, and immune recognition. This knockout model thus enables dissection of how membrane lipid composition influences colorectal cancer phenotypes such as proliferation, migration, and autophagy-dependent survival under metabolic stress. Since SELENOI deficiency is associated with neurodevelopmental disorders and congenital disorders of glycosylation, this cell line provides a tractable system for studying pathological mechanisms related to GPI anchor deficiency outside of neuronal contexts.
Researchers can use the SELENOI Knockout HCT 116 Cell Line in assays such as western blotting and RT-qPCR for gene disruption confirmation, lipidomics for PE profiling, flow cytometry for GPI-anchored proteins, LC3 immunofluorescence for autophagic flux, and MTT viability tests. These applications support colorectal cancer lipid metabolism, GPI anchor biogenesis, and autophagy research. This validated knockout line enables reproducible membrane phospholipid analysis. For more information, contact Ascent Research.
