ESYT2 & ESYT3 Knockout HEK293 Cell Line

The choice depends on whether you are studying combined ESYT2/ESYT3 function or distinguishing ESYT2/ESYT3-dependent from ESYT1-dependent ER-PM contact site functions. The Double Knockout line is uniquely valuable for asking whether ESYT2/ESYT3 are required for these processes — ESYT2 and ESYT3 share substantial functional overlap as ER-PM contact site lipid transfer proteins; single knockouts retain residual activity from the other paralog, while double knockout eliminates ESYT2/ESYT3 functions, leaving only ESYT1. Single-isoform rescue (ESYT2 alone or ESYT3 alone) in the double knockout enables paralog-specific functional dissection. For ER-PM contact site research, the EDITGENE ESYT2 & ESYT3 Double Knockout in HEK293 is the gold-standard genetic tool — combined loss eliminates the most-characterized ESYT family members. Single-isoform rescue (ESYT2 alone or ESYT3 alone) is the gold-standard experimental design for paralog dissection. ESYT1 paralog expression analysis aids interpretation. The double knockout is uniquely valuable for studying glycerophospholipid transfer dynamics, PI(4,5)P2 replenishment at PM, and emerging ESYT family-targeted approaches.

Primary applications: • Complete ESYT2/ESYT3 elimination: ER-PM contact site morphology and lipid transfer analysis — double KO eliminates ESYT2/ESYT3-dependent functions. • Single-isoform rescue: re-introduction of ESYT2 alone or ESYT3 alone enables paralog-specific functional dissection — gold-standard experimental design. • ESYT1 residual analysis: ESYT1 expression analysis to interpret the residual ESYT pool in the double KO. • ER-PM contact site collapse: assessment of contact site frequency and ultrastructure in the combined ESYT2/3-null context. EDITGENE recommends this double knockout as the gold-standard genetic tool for ESYT family-targeted ER-PM contact site research.

Yes, and rescue experiments are uniquely powerful in this double knockout: • Single-isoform rescue: re-introduction of ESYT2 alone or ESYT3 alone in the double knockout enables paralog-specific functional dissection — gold-standard experimental design. • Construct design: use codon-modified ESYT2 or ESYT3 sequences with small C-terminal tags (FLAG, HA). Preserve ER hairpin, SMP domain, and C2 domain architecture. • SMP-domain or C2-domain mutant rescue: enables structure-function dissection within each paralog. • ESYT1 residual: rescue interpretation considers residual ESYT1 expression contributing to ER-PM contact sites. • Functional readout: rescue should restore ER-PM contact site lipid transfer activity. HEK293 transduces efficiently with lentivirus and supports systematic isoform-specific rescue experiments.