Cécile Orsini

MUSEAP, a novel reporter gene for the study of long-term gene expression in immunocompetent mice.

The improvement of gene therapy vectors would benefit from the availability of a reporter gene that can be used for long-term studies in immunocompetent laboratory animals. We describe the construction and characterization of a novel reporter gene, murine secreted embryonic alkaline phosphatase (MUSEAP). We demonstrate by gene transfer in skeletal muscle of immunocompetent mice that MUSEAP is efficiently secreted and detected in the bloodstream and that injection of an increasing dose of DNA leads to a dose-dependent increase of plasma MUSEAP activity. We also show that the expression of MUSEAP under the control of a constitutive promoter is stable for 1 year and that the activity of MUSEAP in the bloodstream reflects the changes in the transcription rate of its gene. These properties make MUSEAP the only reporter gene that can be used for somatic gene transfer into immunocompetent mice in order to study the impact of gene transfer vectors of metabolic, developmental or environmental factors on long-term gene expression.

Transcriptional activation of the metallothionein I gene by electric pulses in vivo: basis for the development of a new gene switch system

In vivo gene transfer to skeletal muscle is a promising strategy for the treatment of muscular disorders and for the systemic delivery of therapeutic proteins. Nevertheless, for a safe and effective protein production, the spatial and temporal control of gene expression is critical. The existing regulating systems rely on the use of an exogenously regulatory protein and/or an inducer drug whose pharmacological properties are of major concerns for therapeutic applications in humans. Therefore, new strategies based on endogenous regulatable elements have been explored.

Comparison of promoter region constructs for in vivo intramuscular expression

High transgene expression is generally expected after gene transfer. However, different level, kinetics and localization of expression might be needed for relevant therapeutic applications. Former studies have compared various promoter regions driving gene expression leading to conflicting results. In the present work, two promoter families have been compared using the efficient in vivo intramuscular electrotransfer technique.

Positive in vivo heterologous gene regulation by electric pulses delivery with metallothionein I gene promoter

In vivo electrotransfer is a physical method of gene delivery in various tissues and organs. It is a promising strategy for the systemic secretion of therapeutic proteins and for DNA vaccination. Nevertheless, for the success of gene therapy in clinics, it is essential to develop gene regulation systems. The existing systems described in the literature all rely on the creation of an artificial transcription factor and/or an inducer drug. New strategies based on endogenous regulatable elements are being developed. We have previously identified the murine metallothionein promoter as an endogenous promoter inducible by controlled electric stimuli applied for electrotransfer experiments. We report here a regulation strategy based on this murine metallothionein promoter in a plasmid context using electric pulses delivery as an inducer.