Tet-On 3G Bidirectional Inducible Expression System (EF1alpha, ZsGreen1)
Tet-On 3G Bidirectional Inducible Expression System (EF1alpha, ZsGreen1)
Brand: Takara Bio.
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Bidirectional Inducible Expression System (EF1alpha, ZsGreen1)
The Tet-On 3G Inducible Expression System (EF1alpha, ZsGreen1) is a powerful, tightly regulated, tetracycline-inducible mammalian expression system that is controlled by the addition of doxycycline to the culture medium. This system allows the simultaneous expression of a gene of interest and a green fluorescent protein (ZsGreen1) marker from a bidirectional, Tet-responsive promoter that binds the Tet-On 3G transactivator protein in the presence of doxycycline. It includes the pTRE3G-BI-ZsGreen1 Vector Set, consisting of a bidirectional vector into which the gene of interest can be cloned and simultaneously expressed with the ZsGreen1 marker, a bidirectional control vector that contains the luciferase gene in one of the two possible cloning sites, and two linear selection markers for hygromycin and puromycin resistance—and the pEF1a-Tet3G Vector, which constitutively expresses the Tet-On 3G transactivator protein from a human elongation factor 1 alpha (EF1α) promoter, allowing expression of the protein in a variety of mammalian cells (such as hematopoietic or stem cells) without the transgene silencing associated with CMV promoters. This system also includes our highly efficient Xfect Transfection Reagent and Tet System Approved FBS.
Overview
- Equivalent co-regulated and simultaneous expression of two genes
- Indirectly monitor expression of a target gene via co-induced expression of a bright green or red fluorescent protein
- Same tight control as all Tet-On 3G systems
- Highly sensitive to doxycycline
- You may alternatively be interested in Tet-On 3G Systems that express two genes from a single IRES bicistronic transcript
Tet-On 3G inducible expression systems with EF1-alpha regulator vectors
Tet-On 3G inducible expression systems with EF1-alpha regulator vectors.
Doxycycline-regulated expression of two genes
Doxycycline-regulated expression of two genes. ZsGreen1 and firefly luciferase were cloned into separate multiple cloning sites that flank the inducible bidirectional promoter in the pTRE3G-BI vector. The plasmid was then cotransfected into HEK 293 cells with pCMV-Tet3G (which expresses the Tet-On 3G transactivator). Cells were treated with the indicated doses of doxycycline, and induced gene expression was measured 24 hr later. Induced expression of ZsGreen1 fluorescent protein was determined by fluorescent microscopy and luciferase activity was measured using a luminometer (RLU = Relative Light Units).
PTRE3G-BI inducible bidirectional promoter shows equivalent induced expression of two genes
PTRE3G-BI inducible bidirectional promoter shows equivalent induced expression of two genes. ZsGreen1 and mCherry fluorescent proteins were cloned into separate multiple cloning sites that flank the inducible bidirectional promoter in the pTRE3G-BI vector. The plasmid was then cotransfected into HEK 293 cells with pEF1a-Tet3G (which expresses the Tet-On 3G transactivator). Cells were treated with 100 ng/ml doxycycline and induced expression of the fluorescent proteins was observed 24 hr later. Cells in the mixed population that expressed high levels of ZsGreen1 also showed similarly high levels of mCherry expression, indicating equivalent expression from both sides of the bidirectional promoter.
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Tet-On 3G inducible expression systems with EF1-alpha regulator vectors
Tet-On 3G inducible expression systems with EF1-alpha regulator vectors.
Doxycycline-regulated expression of two genes
Doxycycline-regulated expression of two genes. ZsGreen1 and firefly luciferase were cloned into separate multiple cloning sites that flank the inducible bidirectional promoter in the pTRE3G-BI vector. The plasmid was then cotransfected into HEK 293 cells with pCMV-Tet3G (which expresses the Tet-On 3G transactivator). Cells were treated with the indicated doses of doxycycline, and induced gene expression was measured 24 hr later. Induced expression of ZsGreen1 fluorescent protein was determined by fluorescent microscopy and luciferase activity was measured using a luminometer (RLU = Relative Light Units).
PTRE3G-BI inducible bidirectional promoter shows equivalent induced expression of two genes
PTRE3G-BI inducible bidirectional promoter shows equivalent induced expression of two genes. ZsGreen1 and mCherry fluorescent proteins were cloned into separate multiple cloning sites that flank the inducible bidirectional promoter in the pTRE3G-BI vector. The plasmid was then cotransfected into HEK 293 cells with pEF1a-Tet3G (which expresses the Tet-On 3G transactivator). Cells were treated with 100 ng/ml doxycycline and induced expression of the fluorescent proteins was observed 24 hr later. Cells in the mixed population that expressed high levels of ZsGreen1 also showed similarly high levels of mCherry expression, indicating equivalent expression from both sides of the bidirectional promoter.
