Vivo-Morpholinos: A Non-Peptide Transporter Delivers Morpholinos into a Wide Array of Mouse Tissues

Morcos, Paul A.; Li, Yongfu; Jiang, Shan

doi:10.2144/000113005

Cited by 189 publications

(206 citation statements)

References 32 publications

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“…All antisense mixtures consist of equal amounts of each antisense oligo. Unmodified morpholinos (PMOs) or octaguanidine dendrimer-conjugated morpholinos (vivo-morpholinos or vPMOs) were used in this study (Gene-Tools) (27,28).…”

Section: Methodsmentioning

confidence: 99%

Bodywide skipping of exons 45–55 in dystrophic mdx52 mice by systemic antisense delivery

Aoki

Yokota

Nagata

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

142

124

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Duchenne muscular dystrophy (DMD), the commonest form of muscular dystrophy, is caused by lack of dystrophin. One of the most promising therapeutic approaches is antisense-mediated elimination of frame-disrupting mutations by exon skipping. However, this approach faces two major hurdles: limited applicability of each individual target exon and uncertain function and stability of each resulting truncated dystrophin. Skipping of exons 45-55 at the mutation hotspot of the DMD gene would address both issues. Theoretically it could rescue more than 60% of patients with deletion mutations. Moreover, spontaneous deletions of this specific region are associated with asymptomatic or exceptionally mild phenotypes. However, such multiple exon skipping of exons 45-55 has proved technically challenging. We have therefore designed antisense oligo (AO) morpholino mixtures to minimize self-or heteroduplex formation. These were tested as conjugates with cell-penetrating moieties (vivo-morpholinos). We have tested the feasibility of skipping exons 45-55 in H2K-mdx52 myotubes and in mdx52 mice, which lack exon 52. Encouragingly, with mixtures of 10 AOs, we demonstrated skipping of all 10 exons in vitro, in H2K-mdx52 myotubes and on intramuscular injection into mdx52 mice. Moreover, in mdx52 mice in vivo, systemic injections of 10 AOs induced extensive dystrophin expression at the subsarcolemma in skeletal muscles throughout the body, producing up to 15% of wild-type dystrophin protein levels, accompanied by improved muscle strength and histopathology without any detectable toxicity. This is a unique successful demonstration of effective rescue by exon 45-55 skipping in a dystrophin-deficient animal model. personalized medicine | nucleic acid therapy | molecular therapy | oligonucleotides | gene therapy D uchenne muscular dystrophy (DMD), the commonest form of muscular dystrophy, is characterized by progressive deterioration of muscle function (1). DMD is caused mainly by frame-shifting deletion or nonsense mutations in the DMD gene, which encodes the protein dystrophin (2). At the milder end of the disease spectrum, Becker muscular dystrophy (BMD) is a form of dystrophin deficiency that presents with a large spectrum of severities, from borderline DMD to almost asymptomatic cases. BMD typically results from in-frame deletions in the DMD gene that allow the expression of limited amounts of an internally truncated but partly functional protein (3).Skipping of exons in DMD muscle so as to restore an in-frame and asymptomatic or very mild Becker-like transcript is among the more promising therapeutic approaches to treatment of DMD (4). To this end, systemic administration of antisense oligonucleotides (AOs) targeting specific exon(s) in the DMD gene has been shown to restore the reading frame and induce body-wide production of partially functional BMD-like dystrophin in mouse and dog models of DMD (5-7). Recently, phase I/II human clinical trials with AOs targeting exon 51 have been completed (8, 9).Although promising, future developm...

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Section: Methodsmentioning

confidence: 99%

Bodywide skipping of exons 45–55 in dystrophic mdx52 mice by systemic antisense delivery

Aoki

Yokota

Nagata

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

142

124

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“…Vivo-Morpholinos, which are octaguanidine-modified antisense oligonucleotides capable of altering pre-mRNA processing in vivo (48), have been used in human cell line-derived xenografts (49). We designed a human GLS pre-mRNA targeting Vivo-Morpholino, which binds to the exon 4-intron 5 junction, blocks junction complex binding to the exon 4 splice donor, and causes exon 4 skipping.…”

Section: Antisense Morpholino Targeting Of Gls Diminishes In Vivo Celmentioning

confidence: 99%

Targeted inhibition of tumor-specific glutaminase diminishes cell-autonomous tumorigenesis

et al. 2015

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“…A new delivery method was recently described where a dendrimer bearing eight guanidinium groups was conjugated to the PMO, which promoted uptake into mammalian cells. 47 PMO conjugates of this design can be administered to mice by intravenous or intraperitoneal injection and penetrate a wide range of tissues; the availability of this new delivery method may enable more widespread use of AMOs made from this interesting chemistry.…”

Section: Inhibiting Mirna Function With Synthetic Oligonucleotides Dementioning

confidence: 99%

Chemical modification and design of anti-miRNA oligonucleotides

2011

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Antisense techniques have been employed for over 30 years to suppress expression of target RNAs. Recently, microRNAs (miRNAs) have emerged as a new class of small, non-coding, regulatory RNA molecules that widely impact gene regulation, differentiation and disease states in both plants and animals. Antisense techniques that employ synthetic oligonucleotides have been used to study miRNA function and some of these compounds may have potential as novel drug candidates to intervene in diseases where miRNAs contribute to the underlying pathophysiology. Anti-miRNA oligonucleotides (AMOs) appear to work primarily through a steric blocking mechanism of action; these compounds are synthetic reverse complements that tightly bind and inactivate the miRNA. A variety of chemical modifications can be used to improve the performance and potency of AMOs. In general, modifications that confer nuclease stability and increase binding affinity improve AMO performance. Chemical modifications and/or certain structural features of the AMO may also facilitate invasion into the miRNA-induced silencing complex. In particular, it is essential that the AMO binds with high affinity to the miRNA 'seed region', which spans bases 2-8 from the 5¢-end of the miRNA.

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Vivo-Morpholinos: A Non-Peptide Transporter Delivers Morpholinos into a Wide Array of Mouse Tissues

Cited by 189 publications

References 32 publications

Bodywide skipping of exons 45–55 in dystrophic mdx52 mice by systemic antisense delivery

Bodywide skipping of exons 45–55 in dystrophic mdx52 mice by systemic antisense delivery

Targeted inhibition of tumor-specific glutaminase diminishes cell-autonomous tumorigenesis

Chemical modification and design of anti-miRNA oligonucleotides

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