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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...

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Autores principales: Asami, Yutaro, Nagata, Tetsuya, Yoshioka, Kotaro, Kunieda, Taiki, Yoshida-Tanaka, Kie, Bennett, C. Frank, Seth, Punit P., Yokota, Takanori
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2021
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.
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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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