TY - JOUR
T1 - Utrophin up-regulation by an artificial transciption factor in transgenic mice
AU - Mattei, Elisabetta
AU - Corbi, Nicoletta
AU - Di Certo, Maria Grazia
AU - Strimpakos, Georgios
AU - Severini, Cinzia
AU - Onori, Annalisa
AU - Desantis, Agata
AU - Libri, Valentina
AU - Buontempo, Serena
AU - Floridi, Aristide
AU - Fanciulli, Maurizio
AU - Baban, Dilair
AU - Davies, Kay E.
AU - Passananti, Claudio
PY - 2007/8/22
Y1 - 2007/8/22
N2 - Duchenne Muscular Dystrophy (DMD) is a severe muscle degenerative disease, due to absence of dystrophin. There is currently, no effective treatment for DMD. Our aim is to up-regulate the expression level of the dystrophin related gene utrophin in DMD, complementing in this way the lack of dystrophin functions. To this end we designed and engineered several synthetic zinc finger based transcription factors. In particular, we have previously shown that the artificial three zinc finger protein named Jazz, fused with the appropriate effector domain, is able to drive the transcription of a test gene from the utrophin promoter "A". Here we report on the characterization of Vp16-Jazz-transgenic mice that specifically over-express the utrophin gene at the muscular level. A Chromatin Immunoprecipitation assay (ChIP) demonstrated the effective access/binding of the Jazz protein to active chromatin in mouse muscle and Vp16-Jazz was shown to be able to up-regulate endogenous utrophin gene expression by immunohistochemistry, western blot analyses and real-time PCR. To our knowledge, this is the first example of a transgenic mouse expressing an artificial gene coding for a zinc finger based transcription factor. The achievement of Vp16-Jazz transgenic mice validates the strategy of transcriptional targeting of endogenous genes and could represent an exclusive animal model for use in drug discovery and therapeutics.
AB - Duchenne Muscular Dystrophy (DMD) is a severe muscle degenerative disease, due to absence of dystrophin. There is currently, no effective treatment for DMD. Our aim is to up-regulate the expression level of the dystrophin related gene utrophin in DMD, complementing in this way the lack of dystrophin functions. To this end we designed and engineered several synthetic zinc finger based transcription factors. In particular, we have previously shown that the artificial three zinc finger protein named Jazz, fused with the appropriate effector domain, is able to drive the transcription of a test gene from the utrophin promoter "A". Here we report on the characterization of Vp16-Jazz-transgenic mice that specifically over-express the utrophin gene at the muscular level. A Chromatin Immunoprecipitation assay (ChIP) demonstrated the effective access/binding of the Jazz protein to active chromatin in mouse muscle and Vp16-Jazz was shown to be able to up-regulate endogenous utrophin gene expression by immunohistochemistry, western blot analyses and real-time PCR. To our knowledge, this is the first example of a transgenic mouse expressing an artificial gene coding for a zinc finger based transcription factor. The achievement of Vp16-Jazz transgenic mice validates the strategy of transcriptional targeting of endogenous genes and could represent an exclusive animal model for use in drug discovery and therapeutics.
UR - http://www.scopus.com/inward/record.url?scp=40749102811&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=40749102811&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0000774
DO - 10.1371/journal.pone.0000774
M3 - Article
C2 - 17712422
AN - SCOPUS:40749102811
VL - 2
JO - PLoS One
JF - PLoS One
SN - 1932-6203
IS - 8
M1 - e774
ER -