Targeting 24 bp within Telomere Repeat Sequences with Tandem Tetramer Pyrrole-Imidazole Polyamide Probes

• 河本, 佑介

  Department of Chemistry, Graduate School of Science, Kyoto University

• 橋谷, かおり

  Structural Biology Center, National Institute of Genetics, and Department of Genetics, School of Life Science, Graduate University for Advanced Studies (Sokendai)

• 板東, 俊和

  Department of Chemistry, Graduate School of Science, Kyoto University

• 杉山, 弘

  Department of Chemistry, Graduate School of Science, Kyoto University

• Ide, Satoru

  Structural Biology Center, National Institute of Genetics, and Department of Genetics, School of Life Science, Graduate University for Advanced Studies (Sokendai)

• Bando, Toshikazu

  Department of Chemistry, Graduate School of Science, Kyoto University

• Maeshima, Kazuhiro

  Structural Biology Center, National Institute of Genetics, and Department of Genetics, School of Life Science, Graduate University for Advanced Studies (Sokendai)

• Sugiyama, Hiroshi

  Department of Chemistry, Graduate School of Science, Kyoto University・Institute for Integrated Cell-Material Science (WPI-iCeMS), Kyoto University

抄録

Synthetic molecules that bind sequence-specifically to DNA have been developed for varied biological applications, including anticancer activity, regulation of gene expression, and visualization of specific genomic regions. Increasing the number of base pairs targeted by synthetic molecules strengthens their sequence specificity. Our group has been working on the development of pyrrole-imidazole polyamides that bind to the minor groove of DNA in a sequence-specific manner without causing denaturation. Recently, we reported a simple synthetic method of fluorescent tandem dimer polyamide probes composed of two hairpin moieties with a linking hinge, which bound to 12 bp in human telomeric repeats (5′-(TTAGGG)n-3′) and could be used to specifically visualize telomeres in chemically fixed cells under mild conditions. We also performed structural optimization and extension of the target base pairs to allow more specific staining of telomeres. In the present study, we synthesized tandem tetramer polyamides composed of four hairpin moieties, targeting 24 bp in telomeric repeats, the longest reported binding site for synthetic, non-nucleic-acid-based, sequence-specific DNA-binding molecules. The novel tandem tetramers bound with a nanomolar dissociation constant to 24 bp sequences made up of four telomeric repeats. Fluorescently labeled tandem tetramer polyamide probes could visualize human telomeres in chemically fixed cells with lower background signals than polyamide probes reported previously, suggesting that they had higher specificity for telomeres. Furthermore, high-throughput sequencing of human genomic DNA pulled down by the biotin-labeled tandem tetramer polyamide probe confirmed its effective binding to telomeric repeats in the complex chromatinized genome.

収録刊行物

• Journal of the American Chemical Society

  Journal of the American Chemical Society 138 (42), 14100-14107, 2016-10-26

  American Chemical Society

詳細情報

• CRID: 1050001335857486336
• NII論文ID: 120006462489
• ISSN: 15205126
• HANDLE: 2433/230915
• 本文言語コード: en
• 資料種別: journal article
• データソース種別:
  • IRDB
  • CiNii Articles