title: Fluorescence imaging of siRNA delivery by peptide nucleic acid-based probe.
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.
authors: Department of Chemistry, Graduate School of Science, Tohoku University.
authors: Takaya
authors: T
authors: Sato
authors: Yusuke
authors: Y
authors: Sato
authors: Kenta
authors: K
authors: Iwai
authors: Shusuke
authors: S
authors: Kuge
authors: Norio
authors: N
authors: Teramae
authors: Seiichi
authors: S
authors: Nishizawa
mesh_terms: D001483:Base Sequence; D052160:Benzothiazoles / Q000737:chemistry; D002470:Cell Survival; D004337:Drug Carriers / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D061848:Optical Imaging / Q000379:methods*; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D011721:Pyrenes / Q000737:chemistry; D011804:Quinolines / Q000737:chemistry; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D052160:Benzothiazoles; D004337:Drug Carriers; D005456:Fluorescent Dyes; D020135:Peptide Nucleic Acids; D011721:Pyrenes; D011804:Quinolines; D034741:RNA, Small Interfering; C051000:thiazole orange; C030984:pyrene