Sequence-selective recognition of double-stranded RNA and enhanced cellular uptake of cationic nucleobase and backbone-modified peptide nucleic acids

Sequence-selective recognition of double-stranded RNA and enhanced cellular uptake of cationic nucleobase and backbone-modified peptide nucleic acids

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 n...

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Bibliographic Details
Published in:RNA (Cambridge) Vol. 23; no. 1; pp. 58 - 69
Main Authors: Hnedzko, Dziyana, McGee, Dennis W, Karamitas, Yannis A, Rozners, Eriks
Format: Journal Article
Language:English
Published: United States Cold Spring Harbor Laboratory Press 01-01-2017
Subjects:
Aminopyridines - metabolism
Arginine - metabolism
Base Sequence
Lysine - metabolism
Models, Molecular
Nucleic Acid Conformation
Peptide Nucleic Acids - metabolism
RNA, Double-Stranded - chemistry
RNA, Double-Stranded - metabolism
cellular uptake of PNA
triple helix
molecular recognition of RNA
modified nucleobases
peptide nucleic acid
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Summary: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 (K = 9.4 to 17 × 10 M ) 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 (K = 0.05 to 1.1 × 10 M ). The binding of double-stranded RNA by M-modified PNA-peptide conjugates was a relatively fast process (k = 2.9 × 10 M sec ) compared to the notoriously slow triple helix formation by oligodeoxynucleotides (k ∼ 10 M sec ). 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.
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content type line 23
ISSN:1355-8382
1469-9001
DOI:10.1261/rna.058362.116