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Transgenic mice expressing an artificial zinc finger regulator targeting an endogenous gene.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2010; Vol. 649, pp. 183-206. - Publication Year :
- 2010
-
Abstract
- 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.
- Subjects :
- Animals
Blotting, Western
Chromatin Immunoprecipitation
Electrophoresis, Polyacrylamide Gel
Female
Genotype
Humans
Immunoblotting
Male
Mice
Mice, Transgenic
Transcription Factors genetics
Utrophin genetics
Genetic Therapy methods
Muscular Dystrophy, Duchenne genetics
Muscular Dystrophy, Duchenne therapy
Transcription Factors metabolism
Zinc Fingers genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 649
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 20680835
- Full Text :
- https://doi.org/10.1007/978-1-60761-753-2_11