SLC25A20 Knockout Huh-7 Cell Line

The choice depends on whether you are studying SLC25A20 (carnitine-acylcarnitine carrier, CAC)'s role in fatty acid β-oxidation or modeling CAC deficiency. The Knockout line is the standard tool for asking whether CAC is required for transporting acylcarnitines across the mitochondrial inner membrane — CAC is essential for fatty acid oxidation by importing acyl-CoA equivalents and exporting free carnitine. Overexpression is useful for testing transport activity or for rescue with disease-associated mutations. For fatty acid metabolism research, the EDITGENE CAC Knockout in Huh-7 is highly relevant — Huh-7 has hepatocellular origin and supports physiologically meaningful fatty acid β-oxidation studies. SLC25A20 mutations cause carnitine-acylcarnitine translocase deficiency, a severe neonatal disorder with hypoketotic hypoglycemia, cardiomyopathy, and high mortality — disease variant rescue enables genotype-function correlation. Rescue with wild-type or transport-deficient CAC is the standard specificity control.

Primary applications: • Fatty acid β-oxidation: oleate or palmitate oxidation assays using ³H- or ¹⁴C-labeled long-chain fatty acids to quantify CAC-dependent fatty acid utilization. • Acylcarnitine profiling: blood-spot or cellular acylcarnitine analysis by tandem mass spectrometry — the standard clinical diagnostic for CAC deficiency. • CAC deficiency modeling: rescue with patient-derived SLC25A20 mutations for genotype-function studies of this severe metabolic disorder. • Drug development: testing potential therapeutic compounds for CAC deficiency in a defined disease-relevant cellular model. EDITGENE recommends this model for researchers investigating fatty acid oxidation, carnitine-acylcarnitine translocase deficiency mechanisms, and pediatric metabolic disease therapeutics.

Yes. CAC rescue experiments are well-established for fatty acid oxidation research: • Construct design: use a codon-modified SLC25A20 sequence with a small C-terminal tag (FLAG, HA). CAC has the SLC25 canonical 6-transmembrane architecture with mitochondrial targeting. • Mitochondrial localization validation: confirm mitochondrial inner membrane localization before fatty acid oxidation assays. • CAC deficiency mutation rescue: patient-derived SLC25A20 mutations enable disease genotype-function correlation studies of this severe neonatal disorder. • Functional readout: rescue should restore fatty acid β-oxidation activity measured by ¹⁴C-palmitate oxidation and normalize acylcarnitine profiles. Huh-7 transduces efficiently with lentivirus and supports stable rescue line generation in a hepatic-relevant context for fatty acid metabolism studies.