ViraPower™ HiPerform™ T-REx™ Gateway™ Expression System
The ViraPower™ HiPerform™ T-REx™ Gateway™ Expression System includes all the components needed to generate lentivirus, including vector kit, 293FT cell line, and the support kit. This kit combines Invitrogen's ViraPower™ HiPerform™ Lentiviral, T-REx™ and Gateway™ technologies to facilitate easy recombination-based cloning and lentiviral-based, regulated (Tetracycline-inducible), high-level expression of a target gene in dividing and non-dividing mammalian cells. The pLenti6.3/ TO/V5-DEST vector is equipped with two key genetic elements, making it a HiPerform™ vector: the Woodchuck Posttranscriptional Regulatory Element (WPRE) and the central Polypurine Tract (cPPT) sequence from the HIV-1 integrase gene to produce at least 4-fold increase in protein expression compared to vectors lacking these elements.
Advantages
- Generates replication-incompetent lentivirus for transducing dividing and non-dividing mammalian cells
- Easy recombination-based cloning using Gateway™ technology
- Stable, long-term, tetracycline-regulated expression
- Enhanced protein expression, up to 4-fold or greater, compared to traditional lentiviral expression systems
Key Features
- WPRE (Woodchuck Posttranscriptional Regulatory Element) from the woodchuck hepatitis virus, increases transgene expression and cPPT (central Polypurine Tract) from the HIV-1 integrase gene, increases the copy number of lentivirus integrating into the host genome, thus increasing viral titer. WPRE and cPPT together produce at least a four-fold increase in protein expression in most cell types, compared to other vectors that do not contain these elements.
- Hybrid promoter consisting of the human cytomegalovirus (CMV) promoter and two tetracycline operator 2 (TetO2) sites for high-level, regulated expression of the target gene
- Blasticidin selection marker for stable selection under control of SV40 promoter
Kit includes
- ViraPower™ HiPerform™ T-Rex™ Gateway™ Vector Kit (Cat. No. A11144)
- ViraPower™ Bsd Lentiviral Support Kit (K497000)
- 293FT Cell Line (R70007)
- Gateway™ LR Clonase™ II Plus Enzyme Mix (12538120)
- One Shot™ Stbl3™ Chemically Competent E. coli (C737303)
- Geneticin™ (10131035)
- pENTR™ Gus positive control plasmid
ViraPower™ HiPerform™ T-Rex™ Gateway™ Vector Kit (Cat. No. A11144)
• pLenti6.3/TO/V5-DEST vector (40 μL of vector at 150 ng/μL in TE Buffer, pH 8.0), 6 μg
• pLenti3.3/TR vector (40 μL of vector at 0.5 μg/μL in TE Buffer, pH 8.0), 20 μg
• pLenti6.3/TO/V5-GW/lacZ plasmid (20 μL of plasmid at 0.5 μg/μL in TE Buffer, pH 8.0), 10 μg
• Tetracycline (10 mg/mL in water), 1 mL. Store at -20°C, protect from light.
Store at -20°C unless otherwise noted
ViraPower™ Bsd Lentiviral Support Kit (Cat. No. K497000)
• ViraPower™ Lentiviral Packaging Mix (Contains 195 μL of a mixture of pLP1, pLP2, and pLP/VSVG plasmids, at 1 μg/μL in TE Buffer, pH 8.0), 195 μg. Store at -20°C.
• Lipofectamine™ 2000 (Cat. No. 11668027), 0.75 mL. Store at 4°C (Do not freeze)
• Blasticidin powder (Cat. No. R21001), 50 mg. Store at -20°C.
293FT Cell Line (Cat. No. R70007)
• 3 x 106 frozen cells in 1 mL of Freezing Medium, 1 vial. Store in liquid nitrogen
Gateway™ LR Clonase™ II Plus Enzyme Mix (Cat. No. 12538120)
• Gateway™ LR Clonase™ II Plus Enzyme Mix, 40 μL. Store -20°C for up to 6 months, -80°C for long term storage.
• Proteinase K Solution (2 ug/mL), 40 μL
One Shot™ Stbl3™ Chemically Competent E. coli (Cat. No. C737303)
• Stbl3™ Cells, 6 mL of S.O.C. Medium, 21 x 50 μL. Store at -80°C.
Geneticin™ (Cat. No. 10131035)
• Geneticin™ solution (50 mg/mL), 20 mL, store at 4°C or -20°C
• pENTR™ Gus positive control plasmid, store at -20°C
pUC19 control DNA, (10 pg/μL), 50 μL