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Fluorescence Imaging of siRNA Delivery by Peptide Nucleic Acid-based Probe

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Abstract

We report on the use of a peptide nucleic acid (PNA)-based fluorescent probe for the analysis of siRNA delivery to living cells. The probe, Py-AA-TO, possesses thiazole orange (TO) and pyrene moieties in the C- and N-termini of PNA, and can function as a light-up probe capable of selective binding to 3′-overhanging nucleotides of target siRNAs. The affinitylabeling of the siRNAs with Py-AA-TO facilitates fluorescence imaging of cellular uptake of polymer-based carriers encapsulating the siRNAs (polyplexes) through endocytosis and subsequent sequestration into lysosome. In addition, flow cytometric measurements reveal that the monitoring of Py-AA-TO fluorescence inside the cells is successfully applicable to the analysis of the polyplex disassembly. These promising functions of Py-AA-TO are presented and discussed as a basis for the design of molecular probes for fluorescent imaging and quantitative analysis of the siRNA delivery process.

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Acknowledgments

We thank Prof. Hidetoshi Oikawa and Dr. Tsunenobu Onodera for measurements of physicochemical parameters, and Prof. Tomokazu Matsue and Dr. Hitoshi Shiku for flow cytometric experiments. This work was supported by Grants-in-Aid for Scientific Research (B) (No. 24350033), Challenging Exploratory Research (No. 25620102) from the Ministry of Education, Culture, Sports, Science and Technology, Japan, and by a grant from the Yamada Science Foundation.

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Authors and Affiliations

  1. Department of Chemistry, Graduate School of Science, Tohoku University, Aoba, Sendai, 980-8578, Japan

    Takaya Sato, Yusuke Sato, Norio Teramae & Seiichi Nishizawa

  2. Department of Microbiology, Tohoku Pharmaceutical University, Aoba, Sendai, 981-8558, Japan

    Kenta Iwai & Shusuke Kuge

Authors
  1. Takaya Sato
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  2. Yusuke Sato
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  3. Kenta Iwai
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  4. Shusuke Kuge
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  5. Norio Teramae
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  6. Seiichi Nishizawa
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Corresponding authors

Correspondence to Yusuke Sato or Seiichi Nishizawa.

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Sato, T., Sato, Y., Iwai, K. et al. Fluorescence Imaging of siRNA Delivery by Peptide Nucleic Acid-based Probe. ANAL. SCI. 31, 315–320 (2015). https://doi.org/10.2116/analsci.31.315

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  • DOI: https://doi.org/10.2116/analsci.31.315

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