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Optimizing antisense oligonucleotides using phosphorodiamidate morpholino oligomers.
Methods Mol Biol. 2012; 867:143-67.MM

Abstract

Duchenne muscular dystrophy (DMD) is caused by mutations that disrupt the reading frame of the human DMD gene. Selective removal of exons flanking an out-of-frame DMD mutation can result in an in-frame mRNA transcript that may be translated into an internally deleted Becker muscular dystrophy-like functionally active dystrophin protein with therapeutic activity. Antisense oligonucleotides (AOs) can be designed to bind to complementary sequences in the targeted mRNA and modify pre-mRNA splicing to correct the reading frame of a mutated transcript. AO-induced exon skipping resulting in functional truncated dystrophin has been demonstrated in animal models of DMD both in vitro and in vivo, in DMD patient cells in vitro in culture, and in DMD muscle explants. The recent advances made in this field suggest that it is likely that AO-induced exon skipping will be the first gene therapy for DMD to reach the clinic. However, it should be noted that personalized molecular medicine may be necessary, since the various reading frame-disrupting mutations are spread across the DMD gene. The different deletions that cause DMD would require skipping of different exons, which would require the optimization and clinical trial workup of many specific AOs. This chapter describes the methodologies available for the optimization of AOs, in particular phosphorodiamidate morpholino oligomers, for the targeted skipping of specific exons on the DMD gene.

Authors+Show Affiliations

School of Biological Sciences, Royal Holloway, University of London, London, UK. linda.popplewell@rhul.ac.ukNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

22454060

Citation

Popplewell, Linda J., et al. "Optimizing Antisense Oligonucleotides Using Phosphorodiamidate Morpholino Oligomers." Methods in Molecular Biology (Clifton, N.J.), vol. 867, 2012, pp. 143-67.
Popplewell LJ, Malerba A, Dickson G. Optimizing antisense oligonucleotides using phosphorodiamidate morpholino oligomers. Methods Mol Biol. 2012;867:143-67.
Popplewell, L. J., Malerba, A., & Dickson, G. (2012). Optimizing antisense oligonucleotides using phosphorodiamidate morpholino oligomers. Methods in Molecular Biology (Clifton, N.J.), 867, 143-67. https://doi.org/10.1007/978-1-61779-767-5_10
Popplewell LJ, Malerba A, Dickson G. Optimizing Antisense Oligonucleotides Using Phosphorodiamidate Morpholino Oligomers. Methods Mol Biol. 2012;867:143-67. PubMed PMID: 22454060.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Optimizing antisense oligonucleotides using phosphorodiamidate morpholino oligomers. AU - Popplewell,Linda J, AU - Malerba,Alberto, AU - Dickson,George, PY - 2012/3/29/entrez PY - 2012/3/29/pubmed PY - 2012/7/24/medline SP - 143 EP - 67 JF - Methods in molecular biology (Clifton, N.J.) JO - Methods Mol Biol VL - 867 N2 - Duchenne muscular dystrophy (DMD) is caused by mutations that disrupt the reading frame of the human DMD gene. Selective removal of exons flanking an out-of-frame DMD mutation can result in an in-frame mRNA transcript that may be translated into an internally deleted Becker muscular dystrophy-like functionally active dystrophin protein with therapeutic activity. Antisense oligonucleotides (AOs) can be designed to bind to complementary sequences in the targeted mRNA and modify pre-mRNA splicing to correct the reading frame of a mutated transcript. AO-induced exon skipping resulting in functional truncated dystrophin has been demonstrated in animal models of DMD both in vitro and in vivo, in DMD patient cells in vitro in culture, and in DMD muscle explants. The recent advances made in this field suggest that it is likely that AO-induced exon skipping will be the first gene therapy for DMD to reach the clinic. However, it should be noted that personalized molecular medicine may be necessary, since the various reading frame-disrupting mutations are spread across the DMD gene. The different deletions that cause DMD would require skipping of different exons, which would require the optimization and clinical trial workup of many specific AOs. This chapter describes the methodologies available for the optimization of AOs, in particular phosphorodiamidate morpholino oligomers, for the targeted skipping of specific exons on the DMD gene. SN - 1940-6029 UR - https://www.unboundmedicine.com/medline/citation/22454060/Optimizing_antisense_oligonucleotides_using_phosphorodiamidate_morpholino_oligomers_ L2 - https://dx.doi.org/10.1007/978-1-61779-767-5_10 DB - PRIME DP - Unbound Medicine ER -