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pTUNE Inducible Vector
pTUNE inducible vector contains a tunable genetic switch that tightly regulates gene expression in mammalian cells. This switch couples repressor proteins and an RNAi target sequence control. More details can be found at Cell 130, 2007.
| 0 µM IPTG |
2.5 µM IPTG |
25 µM IPTG |
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| GFP expression level is controlled by the pTUNE inducible vector in a tunable fashion. HEK293 cells transfected with pTUNE-tGFP treated with different amounts of IPTG. |
Schematic Diagrams of pTUNE inducible vector. In this chart, GFP ORF was used as an example to demonstrate the mechanism of IPTG induction. In TrueORF clones, your gene of interest will be in place of the GFP ORF.
(A) In the off state, repressor proteins (LacI) are expressed and bind to two LacO sites. This results in repression transcription of target gene (GFP is shown as an example in this diagram) and TetR expression. Repression of TetR enables shRNA expression that works on the target sequence located in the 3 ‘-end UTR of the gene, therefore destroy any residual gene expression.
(B) Upon induction with IPTG, the vector switches to on state. IPTG binds to the LacI proteins, resulting its release from LacO sites, which in turn allows the transcription of the target gene and tetR. The latter binds to the tet operator site and shuts down the transcription of the shRNA.
Note: All functional elements, including the shRNA/shRNA target, are all built into the vector. The ORF of interest is the only element that needs to be cloned into the pTUNE vector.
Features:
- Starting from a very tight off-state of >99% repression.
- The gene expression can be fine-tuned as desired.
- The control is reversible.
- Can be easily shuttled from any of OriGene’s TrueORF clones.
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