SLC33A1 Knockout HCT116 Cell Line

The choice depends on whether you are studying SLC33A1 (AT-1)'s role as the principal ER acetyl-CoA transporter or modeling hereditary spastic paraplegia type 42 and congenital cataracts. The Knockout line is the standard tool for asking whether AT-1 is required for delivering acetyl-CoA to the ER lumen for protein acetylation — AT-1 is the only known ER acetyl-CoA importer, essential for substrate supply to ER acetyltransferases like ATase1/ATase2. Overexpression is useful for studying disease-associated AT-1 mutations or for biochemical reconstitution. For ER acetylation research, the EDITGENE SLC33A1 Knockout in HCT 116 is highly informative — AT-1 loss disrupts ER protein quality control, ER stress responses, and reticulophagy. SLC33A1 mutations cause SPG42 (autosomal dominant) and CATIFA syndrome (autosomal recessive) — disease variant rescue enables genotype-function correlation studies. Rescue with wild-type or transport-deficient AT-1 is the standard specificity control.

Primary applications: • ER acetyl-CoA transport: in vitro acetyl-CoA uptake assays using isolated ER microsomes from knockout versus wild-type cells. • ER protein acetylation: analysis of BiP/GRP78, calreticulin, and other ER protein acetylation status by mass spectrometry. • ER stress and reticulophagy: ATF6, IRE1, PERK pathway activation and ER-phagy flux measurement given AT-1's role in ER homeostasis. • SPG42/CATIFA modeling: rescue with disease-associated SLC33A1 mutations for genotype-function studies. EDITGENE recommends this model for researchers investigating ER acetyl-CoA biology, hereditary spastic paraplegia, and ER protein quality control.

Yes. AT-1 rescue experiments require attention to ER membrane targeting: • Construct design: use a codon-modified SLC33A1 sequence with a small C-terminal tag (FLAG, HA). AT-1 has 11 transmembrane domains with ER targeting determinants. • ER localization validation: confirm ER localization by calnexin or PDI co-staining before functional assays. • Disease mutation rescue: SPG42-associated dominant mutations and CATIFA syndrome-associated recessive mutations enable comprehensive disease genotype-function studies. • Functional readout: rescue should restore ER acetyl-CoA transport (in vitro microsomal assays) and ER protein acetylation status (mass spectrometry). HCT 116 transduces efficiently with lentivirus and supports stable rescue line generation.