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


METHODS
Plasmids containing different constructions of the murine metallothionein I (mMT-I) promoter were transfected in mice tibialis-cranalis muscles using the simple skeletal muscle electrotransfer method. The regulation system was studied with the murine secreted alkaline phosphatase (MUSEAP) reporter gene.


RESULTS
The mMT-I promoter can be transiently induced in vivo by application of electric fields. Its inducibility was analyzed in a plasmid context. We demonstrated that the mechanism of this transcriptional induction is not mediated by the cellular entry of metal ions. The ARE (antioxidant-responsive element) sequence was identified as the element responsive to the electric field stimulation.


CONCLUSIONS
This time-control of the expression of a therapeutic gene by physical stimuli could be of value in the context of gene regulation for gene therapy.
Rubenstrunk
Anne
A
UMR 7001 CNRS/ENSCP/Gencell S.A., Centre de Recherche de Vitry-Alfortville, 94403 Vitry Sur Seine, France.
Trollet
Capucine
C
Orsini
Cécile
C
Scherman
Daniel
D
D000469:Alkaline Phosphatase / Q000235:genetics; D000704:Analysis of Variance; D000818:Animals; D003587:Cytomegalovirus; D018274:Electroporation / Q000379:methods*; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D007273:Injections, Intramuscular; D008668:Metallothionein / Q000235:genetics*; D051379:Mice; D009841:Oligonucleotides; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D009841:Oligonucleotides; D008668:Metallothionein; D000469:Alkaline Phosphatase