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Tetracycline-responsive gene expression in mouse brain after amplicon-mediated gene transfer
1F Hoffmann-La Roche Ltd, Basel, Switzerland.
Gene Therapy
|February 12, 1998
Summary
Researchers developed a new amplicon vector for gene transfer, pHermes-tet-lacZ, enabling tetracycline-regulated gene expression in the mouse brain. This advancement offers improved control and flexibility for gene therapy applications.
Area of Science:
- Molecular Biology
- Neuroscience
- Gene Therapy
Background:
- Amplicon vectors utilize Herpes simplex virus (HSV) genetic elements for gene delivery.
- Controlling transgene expression in vivo is crucial for gene therapy applications.
Purpose of the Study:
- To construct and evaluate a novel tetracycline-regulatable amplicon vector, pHermes-tet-lacZ.
- To assess the vector's efficacy and controllability for gene transfer into the mouse brain.
Main Methods:
- Construction of the pHermes-tet-lacZ amplicon vector containing the lacZ gene and tetracycline-responsive elements.
- Propagation of the vector in cell lines (E5, 2-2) with helper viruses (d120, 5dl1.2).
- Stereotactic injection into the mouse hippocampus and assessment of transgene expression and cytotoxicity.
Main Results:
- Successful transgene expression (beta-galactosidase) in the dentate gyrus granular cell layer.
- Demonstrated tetracycline-mediated repression (10-fold) of transgene expression.
- Helper virus influenced the level of repression, indicating tunable control.
Conclusions:
- The pHermes-tet-lacZ amplicon vector provides a flexible and controllable tool for gene transfer in the brain.
- In vivo regulation of transgene expression enhances the utility of amplicon vectors for neuroscience research and potential therapies.
