Long-term and tight control of gene expression in mouse skeletal muscle by a new hybrid human transcription factor

Giuseppe Roscilli, Cira Daniela Rinaudo, Monica Cimino, Elisabetta Sporeno, Stefania Lamartina, Gennaro Ciliberto, Carlo Toniatti

Research output: Contribution to journalArticlepeer-review


Diseases requiring frequent and lifelong injections of recombinant proteins would be more efficaciously treated by intramuscular delivery of genes encoding secretable proteins. However, the success of this approach largely depends on our capability to temporally regulate transcription to regulate transgene expression in muscle. A novel 4-hydroxytamoxifen (4-OHT)-dependent transcriptional regulator (called HEA-3) was constructed by fusing in-frame the DNA binding domain of the human hepatocyte nuclear factor-1α (HNF1α), which is not expressed in muscle cells, a G(521)R mutant of the ligand binding domain of human estrogen receptor-α (ERα), and the activation domain derived from human nuclear factor-ΚB p65 subunit (NF-ΚB p65). We demonstrate that an artificial promoter containing multimeric HNF1α binding sites is silent in muscles and in cell lines that lack endogenous HNF1α. HEA-3 stimulated transcription from this target promoter in a stringent 4-OHT-dependent manner. The dynamic range of transgene regulation was high, because of the low basal activity and high inducibility of the system. Ex vivo, HEA-3 increased expression of the transfected reporter gene by more than 1000-fold in a ligand-dependent manner. In vivo, HEA-3 stimulated by more than 100-fold, the expression of secreted alkaline phosphatases after delivery as plasmid DNA into mouse muscles. Moreover, long-term modulation of the expression of intramuscularly delivered mouse erythropoietin was achieved in immunocompetent mice.

Original languageEnglish
Pages (from-to)653-663
Number of pages11
JournalMolecular Therapy
Issue number5
Publication statusPublished - Nov 1 2002


  • Humanized transcription factor
  • Skeletal muscle
  • Transcriptional regulation

ASJC Scopus subject areas

  • Molecular Biology


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