ACOT8 Knockout Cell Line (Hela)

The choice depends on whether you are studying ACOT8 (acyl-CoA thioesterase 8)'s role as a peroxisomal acyl-CoA thioesterase or modeling its functions in lipid metabolism. The Knockout line is the standard tool for asking whether ACOT8 is required for these processes — ACOT8 is a peroxisomal thioesterase that hydrolyzes acyl-CoA esters (including medium/long-chain, bile acid-CoA, and others) to free fatty acid and CoA, regulating intracellular acyl-CoA/CoA pools and peroxisomal fatty acid oxidation; ACOT8 also has a reported interaction with HIV-1 Nef. Overexpression is useful for studying ACOT8 gain-of-function effects. For peroxisomal lipid metabolism research, the EDITGENE ACOT8 Knockout in HeLa enables study of ACOT8 biology. Other ACOT family members expression analysis aids interpretation. Rescue with wild-type or catalytically-dead ACOT8 enables structure-function studies. The knockout is valuable for studying peroxisomal acyl-CoA thioesterase biology, CoA homeostasis, and peroxisomal fatty acid oxidation regulation.

Primary applications: • Acyl-CoA thioesterase activity: acyl-CoA hydrolysis and CoA pool analysis in ACOT8-null cells. • Peroxisomal fatty acid oxidation: peroxisomal FAO flux analysis given ACOT8's peroxisomal localization. • ACOT family dissection: other ACOT family member expression analysis. • HIV-1 Nef interaction: in heterologous HIV-relevant contexts, ACOT8-Nef interaction studies. EDITGENE recommends this model for researchers investigating peroxisomal acyl-CoA thioesterase biology and CoA homeostasis.

Yes. ACOT8 rescue experiments require attention to peroxisomal targeting: • Construct design: use a codon-modified ACOT8 sequence with a small N-terminal tag (FLAG, HA) — ACOT8 has a C-terminal PTS1 peroxisomal targeting signal that must be preserved for proper localization; C-terminal tags may disrupt peroxisomal import. • Peroxisomal localization validation: confirm peroxisomal localization before functional assays. • Catalytically-dead rescue: active site Ser/Asp/His catalytic triad mutations abolish thioesterase activity. • Functional readout: rescue should restore peroxisomal acyl-CoA thioesterase activity. HeLa transduces efficiently with lentivirus and supports stable rescue line generation.