ESYT3 Knockout Cell Line (HEK293)

The choice depends on whether you are studying ESYT3 (extended synaptotagmin 3)'s role as an ER-plasma membrane contact site protein or its functions in non-vesicular lipid transfer between organelles. The Knockout line is the standard tool for asking whether ESYT3 is required for these processes — extended synaptotagmins (ESYT1, ESYT2, ESYT3) are tethering and lipid transfer proteins at ER-PM contact sites, containing N-terminal hairpin (ER anchor), SMP (synaptotagmin-like mitochondrial-lipid-binding protein) domain for glycerophospholipid transfer, and C-terminal C2 domains (PM binding). Overexpression is useful for studying ESYT3 gain-of-function effects. Important consideration: ESYT family members (ESYT1, ESYT2, ESYT3) share substantial functional overlap — single ESYT3 knockout may show modest phenotypes if ESYT1/2 compensate. This product complements the parallel ESYT2 Knockout and the ESYT2 & ESYT3 Double Knockout in HEK293 (both also available) for systematic ESYT family functional dissection. Rescue with wild-type or lipid-transfer-deficient ESYT3 is the standard specificity control.

Primary applications: • ER-PM contact site morphology: confocal imaging of ER-PM junctions and contact site frequency analysis. • Glycerophospholipid transfer: lipid analytical assays to characterize non-vesicular phospholipid transfer at ER-PM contact sites. • ESYT family dissection: parallel analysis with ESYT2 KO and ESYT2&ESYT3 Double KO in HEK293 (both also available) for paralog-specific characterization. • Calcium signaling: ER-PM contact site-dependent SOCE (store-operated calcium entry) and PI(4,5)P2 replenishment. EDITGENE recommends this model for researchers investigating ER-PM contact site biology, particularly when combined with parallel ESYT2 KO and double KO products.

Yes. ESYT3 rescue experiments require attention to ER-PM contact site architecture: • Construct design: use a codon-modified ESYT3 sequence with a small C-terminal tag (FLAG, HA). ESYT3 has N-terminal hairpin (ER membrane anchor), SMP domain (lipid transfer), and three C2 domains (PM binding) — preserve all elements. • SMP-domain-mutant rescue: SMP domain mutations disrupt lipid transfer activity without affecting tethering. • C2-domain-mutant rescue: C2 domain mutations disrupt PM PI(4,5)P2 binding and ER-PM tethering. • Functional readout: rescue should restore ER-PM contact site formation and glycerophospholipid transfer. HEK293 transduces efficiently with lentivirus and supports stable rescue line generation.