AAVpro Tet-One Inducible Expression System (AAV2)
AAVpro Tet-One Inducible Expression System (AAV2)
Brand: Takara Bio.
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AAVpro Tet-One Inducible Expression System (AAV2)
The AAVpro Tet-One Inducible Expression System is a tetracycline-inducible gene expression system that allows you to produce high titers of recombinant, adeno-associated virus (AAV) for establishing a tightly inducible expression system for your gene of interest in mammalian cells. The all-in-one pAAV-TetOne vector expresses the Tet-On 3G transactivator from the constitutive human PGK promoter, and your gene of interest from the PTRE3GS promoter. In the presence of doxycycline (Dox), the Tet-On 3G transactivator specifically binds and activates high-level transcription from the inducible promoter that controls expression of your gene. There is no selection marker on this vector for mammalian cells. The kit also includes the plasmids for preparation of high-titer AAV particles without the use of a helper virus, and AAV Extraction Solution for isolation of pure AAV particles.
Overview
- All-in-one vector design provides all the power of Tet-On 3G technology in a single vector, making inducible expression as easy as constitutive expression
- Low background and high maximum induced expression
- High sensitivity to doxycycline: near maximum expression at 10 ng/ml doxycycline
- Inducible expression in a broad range of tissues, because the Tet-On 3G transactivator is expressed from the human PGK promoter
- High-titer lentivirus packaging system provided with Lenti-X Tet-One systems
- Retro-X systems incude our Retro-X Universal Packaging System
- The AAVpro system uses a helper-free packaging system and includes extraction reagents for efficient isolation of AAV particles
More Information
Applications
- Tightly controlled inducible expression in mammalian cells
Establishing an inducible expression system in target cells with AAVpro Tet-One
Establishing an inducible expression system in target cells with AAVpro Tet-One. The pAAV-TetOne Vector containing your gene of interest, pRC2-mi342, and pHelper are cotransfected into HEK 293 or HEK 293T cell lines to generate a high-titer AAV particles. The AAV particles are used to transduce your target cells, and Dox is added to express your gene of interest when desired.
Transduction and induction in dividing cell lines with different MOIs of TetOne-Luc Control AAV
Transduction and induction in dividing cell lines with different MOIs of TetOne-Luc Control AAV. 1 x 105 cells were seeded in 12-well plates 12 hr before transduction. Cells were transduced with different MOIs (genomic titer), and 100 ng/ml Dox was added. After 48 hr, cells were harvested and analyzed with a standard firefly luciferase assay using a luminometer.
Induction of luciferase after transduction with TetOne-Luc Control AAV in the presence of different concentrations of Dox
Induction of luciferase after transduction with TetOne-Luc Control AAV in the presence of different concentrations of Dox. 1 x 105 HEK 293 cells were seeded in 12-well plates 12 hr before transduction. Cells were transduced with 1 x 105 MOI (genomic titer), and Dox was added at different concentrations. After 48 hr, cells were harvested and analyzed with a standard firefly luciferase assay using a luminometer.
Different induction times of luciferase after transduction with TetOne-Luc Control AAV
Different induction times of luciferase after transduction with TetOne-Luc Control AAV. 1 x 105 HEK 293 cells were seeded in 12-well plates 12 hr before transduction. Cells were transduced with 1 x 105 MOI (genomic titer), and 100 ng/ml Dox was added. Cells were harvested and analyzed with a standard firefly luciferase assay using a luminometer at various time points after induction.
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Establishing an inducible expression system in target cells with AAVpro Tet-One
Establishing an inducible expression system in target cells with AAVpro Tet-One. The pAAV-TetOne Vector containing your gene of interest, pRC2-mi342, and pHelper are cotransfected into HEK 293 or HEK 293T cell lines to generate a high-titer AAV particles. The AAV particles are used to transduce your target cells, and Dox is added to express your gene of interest when desired.
Transduction and induction in dividing cell lines with different MOIs of TetOne-Luc Control AAV
Transduction and induction in dividing cell lines with different MOIs of TetOne-Luc Control AAV. 1 x 105 cells were seeded in 12-well plates 12 hr before transduction. Cells were transduced with different MOIs (genomic titer), and 100 ng/ml Dox was added. After 48 hr, cells were harvested and analyzed with a standard firefly luciferase assay using a luminometer.
Induction of luciferase after transduction with TetOne-Luc Control AAV in the presence of different concentrations of Dox
Induction of luciferase after transduction with TetOne-Luc Control AAV in the presence of different concentrations of Dox. 1 x 105 HEK 293 cells were seeded in 12-well plates 12 hr before transduction. Cells were transduced with 1 x 105 MOI (genomic titer), and Dox was added at different concentrations. After 48 hr, cells were harvested and analyzed with a standard firefly luciferase assay using a luminometer.
Different induction times of luciferase after transduction with TetOne-Luc Control AAV
Different induction times of luciferase after transduction with TetOne-Luc Control AAV. 1 x 105 HEK 293 cells were seeded in 12-well plates 12 hr before transduction. Cells were transduced with 1 x 105 MOI (genomic titer), and 100 ng/ml Dox was added. Cells were harvested and analyzed with a standard firefly luciferase assay using a luminometer at various time points after induction.
