AAV Mediated Tet-inducible System

AAV inducible gene expression system

Creative BioMart has constructed an AAV inducible gene expression system. We combine the multifunctional AAV vector system with tet-on inducible gene expression system to help you deliver tetracycline-inducible gene expression cassette mediated by AAV in vitro and in vivo.

Technology Description

Tet-on inducible gene expression system is a powerful tool to control the expression time of genes of interest (GOI) in mammalian cells. Our tet-on inducible gene expression vector aims to achieve almost complete silencing of GOI in the absence of tetracycline and its analogues and in the presence of tetracycline or one of its analogues. This is achieved by a multicomponent system that combines active silencing via TTS protein in the absence of tetracycline with strong activation via rtTA protein in the presence of tetracycline. In the absence of tetracycline, the TTS protein derived from the fusion of TetR and krab-ab (transcriptional repressor domain of Kid-1 protein) binds to the tre promoter, leading to active suppression of gene transcription. On the other hand, rtTA protein is derived from the fusion of the mutant Tet repressor and VP16, which binds to tre promoter to activate gene transcription only in the presence of tetracycline.

AAV inducible gene expression system

Workflow Of The System

AAV inducible gene expression system Figure: Diagram of rAAV transduction pathway (Wang, D., 2019)

Firstly, AAV inducible gene expression vector is constructed into E. coli plasmid. It is then transfected into the packaging cells together with the helper plasmid, and the vector region between the two reverse terminal repeats (ITRS) is packaged into the live virus. A tetracycline-inducible gene expression cassette consisting of the tetracycline response element (TRE) promoter driving the user-selected GOI and the ubiquitous or tissue-specific promoter driving the regulatory protein TTS/rtTA is placed between two itrs and introduced into the target cells and other viral genomes. Then gene expression can be turned on in the presence of tetracycline.

Why Choose This System

Switch-like gene activation: our tet-on gene expression vector can be used as a real tetracycline-regulated switch to control gene expression.
High-level expression: tre promoter can drive the high level expression of GOI in its induced state.
Security: AAV is the safest virus vector system at present. AAV is replication-deficient in nature and is unknown to cause any human disease.
The risk of host genome damage is low: AAV vector remains in the nucleus in the form of free DNA after transfection into host cells.
High virus titer: our AAV vector can be packaged into a high titer virus. The titer of AAV virus obtained through our virus packaging service can reach more than 1013 copies of genome per milliliter (GC/ml).
Extensive tropism: packaging our AAV vectors into appropriate serotypes makes it easy to transduce a variety of cell and tissue types from widely used mammalian species, such as human, mouse and rat.
In vitro and in vivo effectiveness: our vectors are usually used to transduce cells in living animals, but they can also be used effectively in vitro.

Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP.

Reference

Wang, D., Tai, P.W.L. & Gao, G. Adeno-associated virus vector as a platform for gene therapy delivery. Nat Rev Drug Discov 18, 358–378 (2019).

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