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aGPCR-HEK: A Stable High-Expression Inducible Mammalian Cell Expression System for Adhesion GPCR Structural Biology Applications - PubMed

7 hours ago
  • #Structural Biology
  • #aGPCR
  • #HEK293S
  • aGPCR-HEK is a stable, tetracycline-inducible mammalian cell expression system for adhesion GPCR (aGPCR) structural biology applications.
  • The protocol involves lentiviral production, transduction, flow cytometry enrichment, suspension culture adaptation, and large-scale induction for cryo-EM analysis.
  • This system yields milligram-scale quantities of correctly folded aGPCR, suitable for structural and biochemical studies.
  • Key features include high-level expression of tagged aGPCRs, efficient genomic integration via lentiviral transduction, and flow cytometry enrichment for enhanced yield.
  • The method is particularly useful for ADGRL4, an aGPCR implicated in tumors, which is challenging to express in conventional insect cell systems.