FADS1 Knockout HAP1 Cell Line

The choice depends on whether you are studying FADS1 (Δ5-desaturase, fatty acid desaturase 1)'s role as a critical enzyme in long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis or modeling FADS1 polymorphism effects on PUFA metabolism. The Knockout line is the standard tool for asking whether FADS1 is required for these processes — FADS1 catalyzes the Δ5-desaturation step in PUFA biosynthesis, converting dihomo-γ-linolenic acid (DGLA, 20:3n-6) to arachidonic acid (AA, 20:4n-6) in the ω-6 pathway and eicosatetraenoic acid (ETA, 20:4n-3) to eicosapentaenoic acid (EPA, 20:5n-3) in the ω-3 pathway. Overexpression is useful for studying FADS1 gain-of-function effects. For lipid metabolism research, the EDITGENE FADS1 Knockout in HAP1 enables study of LC-PUFA biosynthesis. This product complements the parallel FADS2 Knockout in HEK293 (also available) for complete dissection of the Δ5/Δ6-desaturase axis. FADS1 polymorphisms (the FADS1-FADS2 gene cluster on chr 11) are among the strongest genetic predictors of human serum PUFA composition. Rescue with wild-type or catalytically-dead FADS1 enables structure-function studies. The knockout is valuable for studying ω-3/ω-6 PUFA pathway dynamics and FADS polymorphism pharmacogenomic research.

Primary applications: • PUFA biosynthesis: lipid mass spectrometry analysis of ω-3 and ω-6 fatty acids (especially DGLA→AA and ETA→EPA conversions) in FADS1-null cells. • Δ5-desaturase activity: in vitro and cellular Δ5-desaturase activity assays. • FADS1 polymorphism studies: rescue with FADS1 polymorphic variants for pharmacogenomic studies of PUFA metabolism. • Complete FADS1/FADS2 dissection: parallel analysis with FADS2 Knockout in HEK293 (also available) for systematic Δ5/Δ6-desaturase axis studies. EDITGENE recommends this model for researchers investigating LC-PUFA biosynthesis, FADS1-FADS2 cluster pharmacogenomics, and ω-3/ω-6 metabolism.

Yes. FADS1 rescue experiments require attention to ER membrane targeting: • Construct design: use a codon-modified FADS1 sequence with a small C-terminal tag (FLAG, HA). FADS1 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: FADS1 polymorphic variants enable pharmacogenomic studies. • Functional readout: rescue should restore Δ5-desaturation activity measured by ¹³C-DGLA conversion to AA or ETA conversion to EPA. HAP1-specific considerations: • Diploidization: HAP1 cells gradually diploidize during extended culture — confirm ploidy by flow cytometry at the time of phenotypic assay. • Integration site sensitivity: position effects on transgene expression are more pronounced in near-haploid backgrounds; generating multiple independent rescue clones is strongly recommended. • Transduction efficiency: HAP1 transduces with lentivirus at moderate efficiency — increase MOI compared to standard immortalized lines.