2025-05-22 東京科学大学
LNAベースのSSOでは、Singleや相補鎖DNAのHDSSOよりも相補鎖RNAかつ5’/3’末端のみPS修飾のHDSSOでよりエクソンスキッピング活性が高く、mdxマウスでもその効果が見られた。
<関連情報>
- https://www.isct.ac.jp/ja/news/ctjdvdoxm8mv
- https://www.isct.ac.jp/plugins/cms/component_download_file.php?type=2&pageId=&contentsId=1&contentsDataId=1582&prevId=&key=21171f2574ac6af93e7b021f721fe9b1.pdf
- https://www.cell.com/molecular-therapy-family/nucleic-acids/fulltext/S2162-2531(25)00022-8
ヘテロ二重鎖オリゴヌクレオチドのエクソンスキッピング活性に及ぼす化学修飾の影響 Effect of chemical modification on the exon-skipping activity of heteroduplex oligonucleotides
Takenori Shimo ∙ Juri Hasegawa ∙ Kotaro Yoshioka ∙ … ∙ Tetsuya Nagata ∙ Takanori Yokota ∙ Satoshi Obika
Molecular Therapy – Nucleic Acids Published:January 31, 2025
DOI:https://doi.org/10.1016/j.omtn.2025.102468
Abstract
We applied heteroduplex oligonucleotide (HDO) technology, which uses an oligonucleotide hybridized with a complementary strand, to efficiently deliver locked nucleic acid (LNA)-based splice-switching oligonucleotides (SSOs) to the nucleus. Using an in vitro assay involving cationic lipids, we revealed that HDO technology increased the exon-skipping activity of LNA-based SSOs. To assess the effect of heteroduplex SSOs (HDSSOs) on exon-skipping activity, we designed and evaluated various HDSSOs using a series of complementary oligonucleotides with different sugar chemistries (DNA, RNA, and LNA), linkages (phosphodiester; PO and phosphorothioate; PS linkages), and lengths. HDO with different complementary oligonucleotide designs demonstrated a variety of exon-skipping activities. Next, we investigated the intracellular behavior of HDOs, which seemed to affect their efficient exon-skipping activity. We found that HDO technology increased the uptake of both SSOs and complementary oligonucleotides into the nuclei. Additionally, a series of complementary oligonucleotides showed different intracellular stabilities, and complementary oligonucleotide design appears to be one of the key factors affecting efficient exon skipping. Finally, we examined the exon-skipping activity of HDSSOs in mdx mice and found that HDSSOs exhibited higher exon-skipping activity than single-stranded LNA-based SSOs in these mice under intramuscular injections.
