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Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo
We recently reported the antisense properties of a DNA/RNA heteroduplex oligonucleotide consisting of a phosphorothioate DNA-gapmer antisense oligonucleotide (ASO) strand and its complementary phosphodiester RNA/phosphorothioate 2′-O-methyl RNA strand. When α-tocopherol was conjugated with the compl...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7854292/ https://www.ncbi.nlm.nih.gov/pubmed/33290725 http://dx.doi.org/10.1016/j.ymthe.2020.10.017 |
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author | Asami, Yutaro Nagata, Tetsuya Yoshioka, Kotaro Kunieda, Taiki Yoshida-Tanaka, Kie Bennett, C. Frank Seth, Punit P. Yokota, Takanori |
author_facet | Asami, Yutaro Nagata, Tetsuya Yoshioka, Kotaro Kunieda, Taiki Yoshida-Tanaka, Kie Bennett, C. Frank Seth, Punit P. Yokota, Takanori |
author_sort | Asami, Yutaro |
collection | PubMed |
description | We recently reported the antisense properties of a DNA/RNA heteroduplex oligonucleotide consisting of a phosphorothioate DNA-gapmer antisense oligonucleotide (ASO) strand and its complementary phosphodiester RNA/phosphorothioate 2′-O-methyl RNA strand. When α-tocopherol was conjugated with the complementary strand, the heteroduplex oligonucleotide silenced the target RNA more efficiently in vivo than did the parent single-stranded ASO. In this study, we designed a new type of the heteroduplex oligonucleotide, in which the RNA portion of the complementary strand was replaced with phosphodiester DNA, yielding an ASO/DNA double-stranded structure. The ASO/DNA heteroduplex oligonucleotide showed similar activity and liver accumulation as did the original ASO/RNA design. Structure-activity relationship studies of the complementary DNA showed that optimal increases in the potency and the accumulation were seen when the flanks of the phosphodiester DNA complement were protected using 2′-O-methyl RNA and phosphorothioate modifications. Furthermore, evaluation of the degradation kinetics of the complementary strands revealed that the DNA-complementary strand as well as the RNA strand were completely cleaved in vivo. Our results expand the repertoire of chemical modifications that can be used with the heteroduplex oligonucleotide technology, providing greater design flexibility for future therapeutic applications. |
format | Online Article Text |
id | pubmed-7854292 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-78542922022-02-03 Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo Asami, Yutaro Nagata, Tetsuya Yoshioka, Kotaro Kunieda, Taiki Yoshida-Tanaka, Kie Bennett, C. Frank Seth, Punit P. Yokota, Takanori Mol Ther Original Article We recently reported the antisense properties of a DNA/RNA heteroduplex oligonucleotide consisting of a phosphorothioate DNA-gapmer antisense oligonucleotide (ASO) strand and its complementary phosphodiester RNA/phosphorothioate 2′-O-methyl RNA strand. When α-tocopherol was conjugated with the complementary strand, the heteroduplex oligonucleotide silenced the target RNA more efficiently in vivo than did the parent single-stranded ASO. In this study, we designed a new type of the heteroduplex oligonucleotide, in which the RNA portion of the complementary strand was replaced with phosphodiester DNA, yielding an ASO/DNA double-stranded structure. The ASO/DNA heteroduplex oligonucleotide showed similar activity and liver accumulation as did the original ASO/RNA design. Structure-activity relationship studies of the complementary DNA showed that optimal increases in the potency and the accumulation were seen when the flanks of the phosphodiester DNA complement were protected using 2′-O-methyl RNA and phosphorothioate modifications. Furthermore, evaluation of the degradation kinetics of the complementary strands revealed that the DNA-complementary strand as well as the RNA strand were completely cleaved in vivo. Our results expand the repertoire of chemical modifications that can be used with the heteroduplex oligonucleotide technology, providing greater design flexibility for future therapeutic applications. American Society of Gene & Cell Therapy 2021-02-03 2020-12-07 /pmc/articles/PMC7854292/ /pubmed/33290725 http://dx.doi.org/10.1016/j.ymthe.2020.10.017 Text en © 2020 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Asami, Yutaro Nagata, Tetsuya Yoshioka, Kotaro Kunieda, Taiki Yoshida-Tanaka, Kie Bennett, C. Frank Seth, Punit P. Yokota, Takanori Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo |
title | Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo |
title_full | Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo |
title_fullStr | Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo |
title_full_unstemmed | Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo |
title_short | Efficient Gene Suppression by DNA/DNA Double-Stranded Oligonucleotide In Vivo |
title_sort | efficient gene suppression by dna/dna double-stranded oligonucleotide in vivo |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7854292/ https://www.ncbi.nlm.nih.gov/pubmed/33290725 http://dx.doi.org/10.1016/j.ymthe.2020.10.017 |
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