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Exon Skipping and Inclusion Therapies pp 395–411Cite as

Optimization of 2′,4′-BNA/LNA-Based Oligonucleotides for Splicing Modulation In Vitro

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1828))

Abstract

Antisense oligonucleotide-mediated splicing modulation is an attractive strategy for treating genetic disorders. In 2016, two splice-switching oligonucleotides (SSOs) were approved by the FDA. To date, various types of novel artificial nucleic acids have been developed, and their potential for splicing modulations has been demonstrated. To apply these novel chemistries to SSOs, it is necessary to determine the appropriate design for each artificial nucleic acid such as the length of the SSO and number of modifications. In this protocol, we focus on SSOs modified with 2′-O,4′-methylene-bridged nucleic acid (2′,4′-BNA)/locked nucleic acid (LNA), which is an artificial nucleic acid that shows extremely high binding affinity to target RNA strands. We describe our typical protocol for the optimization of 2′,4′-BNA-based SSOs.

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Acknowledgments

T.S. was supported by Grant-in-Aid for JSPS Research Fellow Grant Number 15 J05689.

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Author notes

  1. Takenori Shimo & Satoshi Obika

    Present address: Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, 565-0871, Osaka, Japan

Authors and Affiliations

    Authors
    1. Takenori Shimo
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    2. Satoshi Obika
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    Corresponding author

    Correspondence to Satoshi Obika .

    Editor information

    Editors and Affiliations

    1. Medical Genetics, University of Alberta, Edmonton, AB, Canada

      Toshifumi Yokota

    2. Medical Genetics, University of Alberta, Edmonton, AB, Canada

      Rika Maruyama

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    Shimo, T., Obika, S. (2018). Optimization of 2′,4′-BNA/LNA-Based Oligonucleotides for Splicing Modulation In Vitro. In: Yokota, T., Maruyama, R. (eds) Exon Skipping and Inclusion Therapies. Methods in Molecular Biology, vol 1828. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8651-4_25

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    • DOI: https://doi.org/10.1007/978-1-4939-8651-4_25

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    • Publisher Name: Humana Press, New York, NY

    • Print ISBN: 978-1-4939-8650-7

    • Online ISBN: 978-1-4939-8651-4

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