The Expi293 Inducible GnTI- Expression System Kit is part of the Expi293 platform for rapid, high-yield transient protein expression from mammalian 293 cells. The system kit is centered on engineered Expi293F cells that do not have N-acetylglucosaminyltransferase I (GnTI) activity, therefore lacking complex N-glycans, and stably express the tetracycline repressor (TetR) protein, thus enabling tetracycline-regulated protein expression. The Expi293F Inducible GnTI– cell line was established by the selective knockout of the GnTI gene and the stable integration of the regulatory plasmid pcDNA6/TR, which encodes the TetR gene under the control of the human CMV promoter, in parental Expi293F cells. This cell line is a powerful tool for tetracycline-inducible high-yield expression of homogeneously glycosylated recombinant proteins.
Expi293 Inducible GnTI– Expression System Kit features include: • Homogeneous N-glycosylation of expressed proteins • Tetracycline-inducible protein expression with extremely low basal expression levels • Equivalent protein yields to parental Expi293F cells—up to 1 g/L • pcDNA5/TO expression vector for tetracycline-regulated expression of the gene of interest • Suspension, high-density culture of Expi293F Inducible GnTI– cells in Expi293 Expression Medium • High-efficiency transient transfection using ExpiFectamine 293 reagent in combination with specialized transfection enhancers
The components included in the Expi293 Inducible GnTI– Expression System Kit are: one vial of frozen Expi293F Inducible GnTI– cells, 1 L of Expi293 Expression Medium, one ExpiFectamine 293 transfection kit sufficient to transfect 1 L of culture, 100 mL Opti-Plex Complexation Buffer, one PNGase F Glycan Cleavage kit, one pcDNA5/TO Mammalian Expression Vector kit, and the pRABBIT IgG IRES-EmGFP positive control vector. All components of the Expi293 Inducible GnTI– Expression System are also available for individual purchase.