Transgenic mice expressing an artificial zinc finger regulator targeting an endogenous gene

Methods Mol Biol. 2010:649:183-206. doi: 10.1007/978-1-60761-753-2_11.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Chromatin Immunoprecipitation
  • Electrophoresis, Polyacrylamide Gel
  • Female
  • Genetic Therapy / methods*
  • Genotype
  • Humans
  • Immunoblotting
  • Male
  • Mice
  • Mice, Transgenic
  • Muscular Dystrophy, Duchenne / genetics*
  • Muscular Dystrophy, Duchenne / therapy*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Utrophin / genetics
  • Zinc Fingers / genetics*

Substances

  • Transcription Factors
  • Utrophin