Mesyl phosphoramidate antisense oligonucleotides as an alternative to phosphorothioates with improved biochemical and biological properties
| Autor: | Valentin V. Vlassov, Marina A. Zenkova, O. A. Patutina, Dmitry A. Stetsenko, E. A. Burakova, Boris P. Chelobanov, Sidney Altman, A. A. Fokina, S. K. Miroshnichenko |
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| Rok vydání: | 2019 |
| Předmět: |
Ribonuclease H
Melanoma Experimental Oligonucleotides Apoptosis oncogenic microRNA Biochemistry Phosphates 03 medical and health sciences chemistry.chemical_compound Mice 0302 clinical medicine Cell Movement Cell Line Tumor Animals Phosphoric Acids RNase H 030304 developmental biology Cell Proliferation 0303 health sciences Nuclease Multidisciplinary biology Chemistry Oligonucleotide phosphorothioate DNA Phosphoramidate Biological activity DNA Oligonucleotides Antisense Biological Sciences Amides MicroRNAs 030220 oncology & carcinogenesis Phosphodiester bond biology.protein RNA RNA Cleavage miR-21 |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America |
| ISSN: | 1091-6490 |
| Popis: | Significance Forty years of research have shown that antisense oligonucleotides have great potential to target mRNAs of disease-associated genes and noncoding RNAs. Among the vast number of oligonucleotide backbone modifications, phosphorothioate modification is the most widely used in research and the clinic. However, along with their merits are notable drawbacks of phosphorothioate oligonucleotides, including decreased binding affinity to RNA, reduced specificity, and increased toxicity. Here we report the synthesis and in vitro evaluation of the DNA analog mesyl phosphoramidate oligonucleotide. This oligonucleotide type recruits RNase H and shows significant advantages over phosphorothioate in RNA affinity, nuclease stability, and specificity in inhibiting key processes of carcinogenesis. Thus, mesyl phosphoramidate oligonucleotides may be an attractive alternative to phosphorothioates. Here we describe a DNA analog in which the mesyl (methanesulfonyl) phosphoramidate group is substituted for the natural phosphodiester group at each internucleotidic position. The oligomers show significant advantages over the often-used DNA phosphorothioates in RNA-binding affinity, nuclease stability, and specificity of their antisense action, which involves activation of cellular RNase H enzyme for hybridization-directed RNA cleavage. Biological activity of the oligonucleotide analog was demonstrated with respect to pro-oncogenic miR-21. A 22-nt anti–miR-21 mesyl phosphoramidate oligodeoxynucleotide specifically decreased the miR-21 level in melanoma B16 cells, induced apoptosis, reduced proliferation, and impeded migration of tumor cells, showing superiority over isosequential phosphorothioate oligodeoxynucleotide in the specificity of its biological effect. Lower overall toxicity compared with phosphorothioate and more efficient activation of RNase H are the key advantages of mesyl phosphoramidate oligonucleotides, which may represent a promising group of antisense therapeutic agents. |
| Databáze: | OpenAIRE |
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