FADS2 Knockout Cell Line (HEK293)

The choice depends on whether you are studying FADS2 (Δ6-desaturase, fatty acid desaturase 2)'s role as a critical enzyme in long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis or modeling FADS2 polymorphism effects on PUFA metabolism. The Knockout line is the standard tool for asking whether FADS2 is required for these processes — FADS2 catalyzes the Δ6-desaturation step in PUFA biosynthesis (ALA → SDA in ω-3 pathway; LA → GLA in ω-6 pathway), generating precursors for downstream elongation and Δ5-desaturation (FADS1) to produce EPA, DHA (ω-3) and arachidonic acid (ω-6). Overexpression is useful for studying FADS2 gain-of-function effects. For lipid metabolism research, the EDITGENE FADS2 Knockout in HEK293 enables study of LC-PUFA biosynthesis. FADS2 polymorphisms are associated with serum PUFA levels and ethnic variation in PUFA capacity (Inuit, South Asians have FADS2 variants affecting PUFA metabolism). Rescue with wild-type or catalytically-dead FADS2 enables structure-function studies. The knockout is valuable for studying ω-3/ω-6 PUFA metabolism, dietary PUFA requirements in FADS2-deficient contexts (DHA dietary necessity), and FADS2 polymorphism pharmacogenomic studies.

Primary applications: • PUFA biosynthesis: lipid mass spectrometry analysis of ω-3 (ALA, SDA, EPA, DHA) and ω-6 (LA, GLA, AA) fatty acid levels in FADS2-null versus rescued cells. • Δ6-desaturase activity: in vitro and cellular Δ6-desaturase activity assays. • FADS2 polymorphism studies: rescue with FADS2 polymorphic variants (e.g., rs174575) for pharmacogenomic studies of PUFA metabolism. • DHA dietary requirement: assessment of cellular DHA-dependent processes in FADS2-null cells, where DHA must be obtained from diet. EDITGENE recommends this model for researchers investigating ω-3/ω-6 PUFA metabolism, FADS2 polymorphism pharmacogenomics, and dietary lipid requirement biology.

Yes. FADS2 rescue experiments require attention to ER membrane targeting: • Construct design: use a codon-modified FADS2 sequence with a small C-terminal tag (FLAG, HA). FADS2 has N-terminal cytochrome b5-like domain, central diiron-binding catalytic region, and ER membrane-anchoring hydrophobic regions — preserve all elements. • ER localization validation: confirm endoplasmic reticulum localization before functional assays. • Catalytically-dead rescue: histidine cluster mutations affecting Fe coordination abolish desaturase activity. • Polymorphism rescue: FADS2 polymorphic variants (e.g., rs174575) enable pharmacogenomic studies. • Functional readout: rescue should restore Δ6-desaturation activity measured by ¹³C-ALA conversion to SDA or LA conversion to GLA. HEK293 transduces efficiently with lentivirus and supports stable rescue line generation.