Fluorescence imaging of siRNA delivery by peptide nucleic acid-based probe

Takaya Sato, Yusuke Sato, Kenta Iwai, Shusuke Kuge, Norio Teramae, Seiichi Nishizawa

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)315-320
Number of pages6
Journalanalytical sciences
Volume31
Issue number4
DOIs
Publication statusPublished - 2015

Keywords

  • Affinity-labeling
  • Delivery carrier
  • Fluorescence microscopy
  • Peptide nucleic acid
  • RNA interference (RNAi)
  • SiRNA delivery
  • Small interfering RNA (siRNA)
  • Thiazole orange

ASJC Scopus subject areas

  • Analytical Chemistry

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