Transgenic mice expressing an artificial zinc finger regulator targeting an endogenous gene.
Zinc finger (ZF) proteins belonging to the Cys2-His2 class provide a simple and versatile framework to design novel artificial transcription factors (ATFs) targeted to the desired genes. Our work is based on ZF ATFs engineered to up-regulate the expression level of the dystrophin-related gene utrophin in Duchenne muscular dystrophy (DMD). In particular, on the basis of the "recognition code" that defines specific rules between zinc finger primary structure and potential DNA-binding sites we engineered and selected a new family of artificial transcription factors, whose DNA-binding domain consists in a three zinc finger peptide called "Jazz." Jazz protein binds specifically the 9 bp DNA sequence (5(')-GCT-GCT-GCG-3(')) present in the promoter region of both the human and mouse utrophin gene. We generated a transgenic mouse expressing Jazz protein fused to the strong transcriptional activation domain VP16 and under the control of the muscle specific promoter of the myosin light chain gene. Vp16-Jazz mice display a strong up-regulation of the utrophin at both mRNA and protein levels. To our knowledge, this represents the first example of a transgenic mouse expressing an artificial gene coding for a zinc finger-based transcription factor.
Passananti
Claudio
C
IBPM-CNR, Regina Elena Cancer Institute, Rome, Italy.
Corbi
Nicoletta
N
Onori
Annalisa
A
Di Certo
Maria Grazia
MG
Mattei
Elisabetta
E
D000818:Animals; D015153:Blotting, Western; D047369:Chromatin Immunoprecipitation; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D005838:Genotype; D006801:Humans; D015151:Immunoblotting; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000628:therapy*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D049411:Utrophin / Q000235:genetics; D016335:Zinc Fingers / Q000235:genetics*
D014157:Transcription Factors; D049411:Utrophin