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

Research paper by Takaya T Sato, Yusuke Y Sato, Kenta K Iwai, Shusuke S Kuge, Norio N Teramae, Seiichi S Nishizawa

Indexed on: 14 Apr '15Published on: 14 Apr '15Published in: Analytical sciences : the international journal of the Japan Society for Analytical Chemistry



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 affinity-labeling 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.