Development or order-mede type artificial restriction enzymes and artificial repressers

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In order to develop therapeutic strategiea through specific modulation of the transcription of target genes, We studied regulation of the transcription level by artificial repressors. In addition, artificial restriction enzymes were designed as useful tool of molecular biology. Many transcription factors are also known to induce DNA bending Therefore, the protein-induced DNA bending is helpful for many combination of protein-protein and/or protein-DNA interactions that are necessary for various biological reactions. The zinc finger motif of Cys_2His_2-type is one of the most common DNA binding motifs. Therefore, this zinc finger motif offers anatractive framework for the design of novel DNA binding proteins. On the basis of the DNA recognition mode and the structural features unique to the Cys_2His_2 zinc finger of DNA binding, approaches to link this zinc fingers with other functional modules such as DNA binding domains to generate artificial chimeric peptides with long binding sitea and DNA-cleavage modules to produce novel sequence specific nucleases, were performed. Indeed, the artificial chimeric DNA binding peptides displayed sequence specificity for extended, chimeric DNA binding sites. Our designed nine-zinc finger peptides also bound 18 or 27 contiguous base pairs of DNA in a sequence-specific fashion. Furthermore, we created six zinc finger proteins by connecting two DNA binding domains through flexible polyglycine linkers. The new proteins induced DNA bending at the intervening region of the two distal binding sites. In summary, artificial zinc finger proteins could find broad application in future gene therapy strategies.