VPS13A Knockout Cell Line (HEK293)

The choice depends on whether you are studying VPS13A's role in lipid transfer at membrane contact sites or modeling chorea-acanthocytosis. The Knockout line is the standard tool for the latter — chorea-acanthocytosis is caused by loss-of-function VPS13A mutations, so KO recapitulates the disease genetics. Overexpression of full-length VPS13A is technically challenging due to its very large size (~3,200 amino acids), making KO the more practically tractable approach for most research questions. For VPS13A research, the EDITGENE Knockout line in HEK293 is the more practically useful model. Rescue experiments often use truncated VPS13A constructs (lipid transfer domain alone, or specific functional fragments) rather than full-length protein, which is particularly informative for mapping domain-specific functions in lipid transfer between organelles.

Primary applications: • Membrane contact site studies: imaging-based analysis of ER-mitochondria, ER-lipid droplet, and ER-endosome contact sites following VPS13A loss. • Lipid composition analysis: lipidomic profiling of membrane fractions to identify lipid transfer dependencies on VPS13A. • Autophagy and lipophagy: LC3, p62, and lipid droplet dynamics assays to assess VPS13A's role in autophagy-related processes. • Erythrocyte morphology (if relevant): for chorea-acanthocytosis modeling, though HEK293 is not the disease-relevant cell type. EDITGENE recommends this model for researchers investigating lipid transfer at membrane contact sites, chorea-acanthocytosis biology, and organelle communication.

Yes, with important caveats due to VPS13A's exceptional size: • Construct design: full-length VPS13A is ~3,200 amino acids and difficult to clone and express in standard vectors. Many published rescue studies use lentiviral or BAC-based delivery systems specifically optimized for large transgenes. • Truncation rescue: lipid transfer domain alone, or specific functional fragments, are commonly used for domain mapping studies and are technically more tractable than full-length rescue. • Tag placement: C-terminal tags are generally preferred; the N-terminus contains protein interaction motifs that should be preserved. • Functional readout: rescue should restore membrane contact site distributions and lipid composition profiles. Full functional rescue with truncation constructs is rarely achieved due to multiple protein interaction interfaces along the length of VPS13A. HEK293 transduces efficiently with lentivirus, though large cargo size reduces titer; consider multi-vector strategies or recombinant adenovirus for full-length VPS13A delivery.