The artificial zinc finger coding gene 'Jazz' binds the utrophin promoter and activates transcription

Up-regulation of utrophin gene expression is recognized as a plausible ther apeutic approach in the treatment of Duchenne muscular dystrophy (DMD). We have designed and engineered new zinc finger-based transcription factors ca pable of binding and activating transcription from the promoter of the dyst rophin-related gene, utrophin. Using the recognition 'code' that proposes s pecific rules between zinc finger primary structure and potential DNA bindi ng sites, we engineered a new gene named 'Jazz' that encodes for a three-zi nc finger peptide. Jazz belongs to the Cys(2)-His(2) zinc finger type and w as engineered to target the nine base pair DNA sequence: 5'-GCT-GCT-GCG-3', present in the promoter region of both the human and mouse utrophin gene. The entire zinc finger cu-helix region, containing the amino acid positions that are crucial for DNA binding, was specifically chosen on the basis of the contacts more frequently represented in the available list of the 'code '. Here we demonstrate that Jazz protein binds specifically to the double-s tranded DNA target, with a dissociation constant of about 32 nM. Band shift and super-shift experiments confirmed the high affinity and specificity of Jazz protein for its DNA target. Moreover, we show that chimeric proteins, named Gal4-Jazz and Sp1-Jazz, are able to drive the transcription of a tes t gene from the human utrophin promoter.