2002 Fiscal Year Final Research Report Summary
Development or order-mede type artificial restriction enzymes and artificial repressers
| Project/Area Number |
12793008
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| Research Category |
Grant-in-Aid for University and Society Collaboration
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| Allocation Type | Single-year Grants |
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| Research Field |
Chemical pharmacy
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
SUGIURA Yukio Kyoto Univ., Inst. Chem. Res., Prof., 化学研究所, 教授 (40025698)
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| Co-Investigator(Kenkyū-buntansha) |
NAGAOKA Makoto Kyoto Univ., Inst. Chem. Res., Assistant Prof., 化学研究所, 助手 (60314275)
GOTOH Susumu Kyoto Univ., Inst. Chem. Res., Associate Prof., 化学研究所, 助教授 (40263149)
FUTAKI Shiroh Kyoto Univ., Inst. Chem. Res., Associate Prof., 化学研究所, 助教授 (50199402)
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| Project Period (FY) |
2000 – 2002
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| Keywords | Zinc finger / Artificial protein / DNA recognition / DNA motif / DNA bending / Gene target / Artificial restriction enzyme / Artificial repressor |
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| Research Abstract |
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.
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