title: Synthesis and properties of peptide nucleic acid labeled at the N-terminus with HiLyte Fluor 488 fluorescent dye.
abstract: Fluorescently labeled peptide nucleic acids (PNAs) are important tools in fundamental research and biomedical applications. However, synthesis of labeled PNAs, especially using modern and expensive dyes, is less explored than similar preparations of oligonucleotide dye conjugates. Herein, we present a simple procedure for labeling of the PNA N-terminus with HiLyte Fluor 488 as the last step of solid phase PNA synthesis. A minimum excess of 1.25equiv of activated carboxylic acid achieved labeling yields close to 90% providing a good compromise between the price of dye and the yield of product and significant improvement over previous literature procedures. The HiLyte Fluor 488-labeled PNAs retained the RNA binding ability and in live cell fluorescence microscopy experiments were brighter and significantly more photostable than PNA labeled with carboxyfluorescein. In contrast to fluorescein-labeled PNA, the fluorescence of PNAs labeled with HiLyte Fluor 488 was independent of pH in the biologically relevant range of 5-8. The potential of HiLyte Fluor 488-labeling for studies of PNA cellular uptake and distribution was demonstrated in several cell lines.
authors.affiliation: Department of Chemistry, Binghamton University, The State University of New York, Binghamton, NY 13902, United States.
authors.forename: Dziyana
authors.initials: D
authors.lastname: Hnedzko
authors.affiliation: Department of Biological Sciences, Binghamton University, The State University of New York, Binghamton, NY 13902, United States.
authors.forename: Dennis W
authors.initials: DW
authors.lastname: McGee
authors.affiliation: Department of Chemistry, Binghamton University, The State University of New York, Binghamton, NY 13902, United States. Electronic address: erozners@binghamton.edu.
authors.forename: Eriks
authors.initials: E
authors.lastname: Rozners
mesh_terms: D045744:Cell Line, Tumor; D005434:Flow Cytometry; D005452:Fluoresceins / Q000737:chemistry / Q000528:radiation effects; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000528:radiation effects; D057809:HEK293 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008856:Microscopy, Fluorescence; D020135:Peptide Nucleic Acids / Q000138:chemical synthesis / Q000737:chemistry* / Q000528:radiation effects; D038761:Photobleaching; D012330:RNA, Double-Stranded / Q000737:chemistry; D012235:Rhodamines / Q000737:chemistry* / Q000528:radiation effects
chemical_list: D005452:Fluoresceins; D005456:Fluorescent Dyes; D020135:Peptide Nucleic Acids; D012330:RNA, Double-Stranded; D012235:Rhodamines; C024098:6-carboxyfluorescein
keywords: Fluorescence microscopy; Fluorescent probes; HiLyte Fluor 488; Peptide nucleic acids; RNA recognition