title: Sequence-selective recognition of double-stranded RNA and enhanced cellular uptake of cationic nucleobase and backbone-modified peptide nucleic acids.
abstract: Sequence-selective recognition of complex RNAs in live cells could find broad applications in biology, biomedical research, and biotechnology. However, specific recognition of structured RNA is challenging, and generally applicable and effective methods are lacking. Recently, we found that peptide nucleic acids (PNAs) were unusually well-suited ligands for recognition of double-stranded RNAs. Herein, we report that 2-aminopyridine (M) modified PNAs and their conjugates with lysine and arginine tripeptides form strong (Ka= 9.4 to 17 × 107M-1) and sequence-selective triple helices with RNA hairpins at physiological pH and salt concentration. The affinity of PNA-peptide conjugates for the matched RNA hairpins was unusually high compared to the much lower affinity for DNA hairpins of the same sequence (Ka= 0.05 to 1.1 × 107M-1). The binding of double-stranded RNA by M-modified PNA-peptide conjugates was a relatively fast process (kon= 2.9 × 104M-1sec-1) compared to the notoriously slow triple helix formation by oligodeoxynucleotides (kon∼ 103M-1sec-1). M-modified PNA-peptide conjugates were not cytotoxic and were efficiently delivered in the cytosol of HEK293 cells at 10 µM. Surprisingly, M-modified PNAs without peptide conjugation were also taken up by HEK293 cells, which, to the best of our knowledge, is the first example of heterocyclic base modification that enhances the cellular uptake of PNA. Our results suggest that M-modified PNA-peptide conjugates are promising probes for sequence-selective recognition of double-stranded RNA in live cells and other biological systems.
authors.affiliation: Department of Chemistry, Binghamton University, State University of New York, Binghamton, New York 13902, USA.
authors.forename: Dziyana
authors.initials: D
authors.lastname: Hnedzko
authors.affiliation: Department of Biological Sciences, Binghamton University, State University of New York, Binghamton, New York 13902, USA.
authors.forename: Dennis W
authors.initials: DW
authors.lastname: McGee
authors.affiliation: Department of Chemistry, Binghamton University, State University of New York, Binghamton, New York 13902, USA.
authors.forename: Yannis A
authors.initials: YA
authors.lastname: Karamitas
authors.affiliation: Department of Chemistry, Binghamton University, State University of New York, Binghamton, New York 13902, USA.
authors.forename: Eriks
authors.initials: E
authors.lastname: Rozners
mesh_terms: D000631:Aminopyridines / Q000378:metabolism; D001120:Arginine / Q000378:metabolism; D001483:Base Sequence; D008239:Lysine / Q000378:metabolism; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D020135:Peptide Nucleic Acids / Q000378:metabolism*; D012330:RNA, Double-Stranded / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000631:Aminopyridines; D020135:Peptide Nucleic Acids; D012330:RNA, Double-Stranded; D001120:Arginine; D008239:Lysine; C032439:alpha-aminopyridine
keywords: cellular uptake of PNA; modified nucleobases; molecular recognition of RNA; peptide nucleic acid; triple helix