We constructed a mouse model over-expressing in muscle the artificial transcription factor "Jazz" showing up-regulation of the endogenous dystrophin related gene "utrophin". Overexpression of utrophin gene represents one of the main therapeutic strategies proposed for Duchenne and Becker Dystrophies. In our knowledge, the Jazz-transgenic model is the first "canonical" transgenic-mouse over-expressing a synthetic zinc finger transcription factor (ZFP TF). We engineered the Jazz family of ZFP TFs, whose DNA binding domain consists in a three-zinc finger peptide. Jazz binds specifically the 9 base pairs DNA sequence present in the promoter region of both the human and mouse utrophin genes. We showed that the chimeric proteins obtained fusing Jazz with the appropriate effector are able to drive the transcription from the utrophin promoter and to bind the target endogenous chromosomal site. To generate muscle specific Jazz-transgenic mice we engineered a construct containing the Myosin Light Chain promoter region, and Jazz fused with the strong activation domain Vp16. We obtained several positive founders (F0) that appear viable, healthy and fertile. On F1 positive mice, western blot analysis of muscle tissues revealed different levels of Vp16-Jazz expression, from moderate to very high. In particular the progeny derived from founders 9 and 41 display an up-regulation of Utrophin, with the interesting appearance of additional utrophin isoforms. The Jazz-transgenic represents an appealing animal model system for applications in basic science, biotechnology, molecular medicine and gene therapy.
TRANSGENIC MOUSE EXPRESSING A SYNTHETIC ZINC FINGER-BASED TRANSCRIPTION FACTOR FOR UTROPHIN REGULATION. A POSSIBLE MODEL FOR PREVENTION AND THERAPY OF MUSCLE DYSTROPHIES
MGDi Certo;A Onori;
2005
Abstract
We constructed a mouse model over-expressing in muscle the artificial transcription factor "Jazz" showing up-regulation of the endogenous dystrophin related gene "utrophin". Overexpression of utrophin gene represents one of the main therapeutic strategies proposed for Duchenne and Becker Dystrophies. In our knowledge, the Jazz-transgenic model is the first "canonical" transgenic-mouse over-expressing a synthetic zinc finger transcription factor (ZFP TF). We engineered the Jazz family of ZFP TFs, whose DNA binding domain consists in a three-zinc finger peptide. Jazz binds specifically the 9 base pairs DNA sequence present in the promoter region of both the human and mouse utrophin genes. We showed that the chimeric proteins obtained fusing Jazz with the appropriate effector are able to drive the transcription from the utrophin promoter and to bind the target endogenous chromosomal site. To generate muscle specific Jazz-transgenic mice we engineered a construct containing the Myosin Light Chain promoter region, and Jazz fused with the strong activation domain Vp16. We obtained several positive founders (F0) that appear viable, healthy and fertile. On F1 positive mice, western blot analysis of muscle tissues revealed different levels of Vp16-Jazz expression, from moderate to very high. In particular the progeny derived from founders 9 and 41 display an up-regulation of Utrophin, with the interesting appearance of additional utrophin isoforms. The Jazz-transgenic represents an appealing animal model system for applications in basic science, biotechnology, molecular medicine and gene therapy.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
