sequence of a eukaryotic gene's DNA that is not translated into a protein
Sequences were
designed to target exonic sites of exon 6 (Ex6A) and exon 8
(Ex8A), or exon/intron boundary between exon 6 and intron
6 (Ex6B), or exon 8 and intron 8 (Ex8B) (see Fig 1).
sequence of a gene's DNA that transcribes into protein structures
A method to rescue dystrophin production by antisense oligonucleotides, termed exon-skipping, has been reported for
the mdx mouse and in four DMD patients by local intramuscular injection.
Most DMD mutations are caused by outof-
frame (frameshift) or nonsense gene mutations,
whereas the majority of BMD mutations are in-frame,
and thus compatible with production of a messenger
RNA (mRNA) transcript that can be translated into a
partly functional quasi-dystrophin (reading frame
rule).3
A method to rescue dystrophin production by antisense oligonucleotides, termed exon-skipping, has been reported for
the mdx mouse and in four DMD patients by local intramuscular injection.
a mutation due to an intramolecular reorganization of a gene
The canine X-linked muscular dystrophy (CXMD)
harbors a point mutation within the acceptor splice site
of exon 7, leading to exclusion of exon 7 from the
mRNA transcript.9
Methods: We tested a series of antisense drugs singly and as cocktails, both in primary cell culture, and two in vivo delivery
methods (intramuscular injection and systemic intravenous injection).
observable characteristics produced by genes and environment
This raises the possibility of using
antisense-mediated removal of exons carrying nonsense
mutations, or whose presence disrupts the open reading
frame at the site of the mutation, so as to restore the
translational reading frame and thus to convert DMD
to a milder BMD phenotype.5
affecting an entire structure, network, or complex of parts
Methods: We tested a series of antisense drugs singly and as cocktails, both in primary cell culture, and two in vivo delivery
methods (intramuscular injection and systemic intravenous injection).
of or relating to uniform movement without rotation
This raises the possibility of using
antisense-mediated removal of exons carrying nonsense
mutations, or whose presence disrupts the open reading
frame at the site of the mutation, so as to restore the
translational reading frame and thus to convert DMD
to a milder BMD phenotype.5
a mutation due to an intramolecular reorganization of a gene
Most DMD mutations are caused by outof-
frame (frameshift) or nonsense gene mutations,
whereas the majority of BMD mutations are in-frame,
and thus compatible with production of a messenger
RNA (mRNA) transcript that can be translated into a
partly functional quasi-dystrophin (reading frame
rule).3
an electric cell that generates an electromotive force by an irreversible conversion of chemical to electrical energy; cannot be recharged
Methods: We tested a series of antisense drugs singly and as cocktails, both in primary cell culture, and two in vivo delivery
methods (intramuscular injection and systemic intravenous injection).
an enzyme that catalyzes the conversion of a proenzyme to an active enzyme
Some BMD patients with deletions as large as
33 exons (46% of the gene) can show little or no clinical
symptoms, with only increased serum creatine kinase
concentration.4
Ann Neurol 2009;65:667–676
Duchenne muscular dystrophy (DMD) and its milder
form, Becker muscular dystrophy (BMD), are caused
by mutations in the DMD gene.1
the branch of medicine that deals with the nervous system
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
the scientific study of the nervous system and the brain
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
affecting or characteristic of both nerves and muscles
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
the capacity to be useful or serve a particular purpose well
Use of cocktails of morpholino, as
shown here, allows broader application of this approach to a greater proportion of DMD patients (90%) and also offers the
prospect of selecting deletions that optimize the functionality of the dystrophin protein.
thickening of tissue in the motor tracts of the lateral columns and anterior horns of the spinal cord; results in progressive muscle atrophy that starts in the limbs
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
an amino acid that does not occur in proteins but is found in the muscle tissue of vertebrates both in the free form and as phosphocreatine; supplies energy for muscle contraction
Some BMD patients with deletions as large as
33 exons (46% of the gene) can show little or no clinical
symptoms, with only increased serum creatine kinase
concentration.4
the use of nuclear magnetic resonance of protons to produce proton density images
This was
accompanied by reduced inflammatory signals examined by magnetic resonance imaging and histology, improved or stabilized
timed running tests, and clinical symptoms.
resonance of electrons or atoms or molecules or nuclei to radiation frequencies as a result of space quantization in a magnetic field
This was
accompanied by reduced inflammatory signals examined by magnetic resonance imaging and histology, improved or stabilized
timed running tests, and clinical symptoms.
the person (or institution) who accepts a check or draft and becomes responsible for paying the party named in the draft when it matures
The canine X-linked muscular dystrophy (CXMD)
harbors a point mutation within the acceptor splice site
of exon 7, leading to exclusion of exon 7 from the
mRNA transcript.9
the forceful insertion of a substance under pressure
A method to rescue dystrophin production by antisense oligonucleotides, termed exon-skipping, has been reported for
the mdx mouse and in four DMD patients by local intramuscular injection.
This was
accompanied by reduced inflammatory signals examined by magnetic resonance imaging and histology, improved or stabilized
timed running tests, and clinical symptoms.
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
in an artificial environment outside the living organism
Previously,
McClorey and colleagues10 showed transfection
with antisense oligo targeting exons 6 and 8 restored
reading frame of mRNA in cultured myotubes from
dystrophic dogs in vitro.
Sequences were
synthesized using two different backbone chemistries: 2OMePs
(Eurogentec Lie`ge, Belgium), and morpholino
(Gene-Tools, LLC Philomath, OR).12
The canine X-linked muscular dystrophy (CXMD)
harbors a point mutation within the acceptor splice site
of exon 7, leading to exclusion of exon 7 from the
mRNA transcript.9
any pathological hardening or thickening of tissue
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
the act of providing treatment for an illness or disorder
Interpretation: This is the first report of widespread rescue of dystrophin expression to therapeutic levels in the dog model of
DMD. This study also provides a proof of concept for systemic multiexon-skipping therapy.
relating to the simplest units of an element or compound
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
part of DNA controlling physical characteristics and growth
Ann Neurol 2009;65:667–676
Duchenne muscular dystrophy (DMD) and its milder
form, Becker muscular dystrophy (BMD), are caused
by mutations in the DMD gene.1
the ability to form mental images of things or events
This was
accompanied by reduced inflammatory signals examined by magnetic resonance imaging and histology, improved or stabilized
timed running tests, and clinical symptoms.
Most DMD mutations are caused by outof-
frame (frameshift) or nonsense gene mutations,
whereas the majority of BMD mutations are in-frame,
and thus compatible with production of a messenger
RNA (mRNA) transcript that can be translated into a
partly functional quasi-dystrophin (reading frame
rule).3
relating to the study of heredity and variation in organisms
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
(computer science) a system of world-wide electronic communication in which a computer user can compose a message at one terminal that can be regenerated at the recipient's terminal when the recipient logs in
E-mail: [email protected] or Dr. Takeda, Department of
Molecular Therapy National Institute of Neuroscience, National
Center of Neurology and Psychiatry 4-1-1 Ogawa-higashi, Kodaiva,
Tokyo, Japan.
act between parties with a view to reconciling differences
This raises the possibility of using
antisense-mediated removal of exons carrying nonsense
mutations, or whose presence disrupts the open reading
frame at the site of the mutation, so as to restore the
translational reading frame and thus to convert DMD
to a milder BMD phenotype.5
This was
accompanied by reduced inflammatory signals examined by magnetic resonance imaging and histology, improved or stabilized
timed running tests, and clinical symptoms.
a dog, wolf, jackal, or other closely related mammal
The canine X-linked muscular dystrophy (CXMD)
harbors a point mutation within the acceptor splice site
of exon 7, leading to exclusion of exon 7 from the
mRNA transcript.9
Antisense Sequences and Chemistries
We designed four antisense sequences to target exons 6
and 8 of the dog dystrophin mRNA as follows: Ex6A
(GTTGATTGTCGGACCCAGCTCAGG), Ex6B (ACCTATGACTGTGGATGAGAGCGTT),
Ex8A (CTTCCTGGATGGCTTCAATGCTCAC),
and Ex8B (ACCTGTTGAGAATAGTGCATTTGAT).
This was
accompanied by reduced inflammatory signals examined by magnetic resonance imaging and histology, improved or stabilized
timed running tests, and clinical symptoms.
an amber, watery fluid, rich in proteins, that separates out when blood coagulates
Some BMD patients with deletions as large as
33 exons (46% of the gene) can show little or no clinical
symptoms, with only increased serum creatine kinase
concentration.4
Here, we identified a phosphorodiamidate
morpholino oligomer (PMO) cocktail
that, using either intramuscular injection or systemic
intravenous delivery, was not toxic, resulted in extensive
dystrophin expression to therapeutic levels, and
was associated with significant functional stabilization
in dystrophic dogs in vivo.
able to exist and perform in harmonious combination
Most DMD mutations are caused by outof-
frame (frameshift) or nonsense gene mutations,
whereas the majority of BMD mutations are in-frame,
and thus compatible with production of a messenger
RNA (mRNA) transcript that can be translated into a
partly functional quasi-dystrophin (reading frame
rule).3
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
Address correspondence to Dr Hoffman, Research Center for Genetic
Medicine, Children’s National Medical Center, Department of
Integrative Systems Biology, George Washington University School
of Medicine, 111 Michigan Avenue NW, Washington, DC 20010.
the series of vertebrae forming the axis of the skeleton and protecting the spinal cord
Sequences were
synthesized using two different backbone chemistries: 2OMePs
(Eurogentec Lie`ge, Belgium), and morpholino
(Gene-Tools, LLC Philomath, OR).12
the profession devoted to alleviating diseases and injuries
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
The ability to use cocktails of antisense
oligonucleotides targeting multiple exons would
permit design of quasi-dystrophin proteins that retain
more functionality,7 Finally, the use of cocktails could
lead to FDA-approved mixtures that would successfully
treat a large group of DMD patients with distinct but
overlapping deletions.
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
of or relating to or caused by attraction for iron
This was
accompanied by reduced inflammatory signals examined by magnetic resonance imaging and histology, improved or stabilized
timed running tests, and clinical symptoms.
a workplace for the conduct of scientific research
This might alleviate the problem
From the 1Research Center for Genetic Medicine, Children’s National
Medical Center, Washington, DC; 2McColl-Lockwood Laboratory
for Muscular Dystrophy Research, Neuromuscular/Amyotrophic
Lateral Sclerosis Center, Carolinas Medical Center,
Charlotte, NC; and 3Department of Molecular Therapy, National
Institute of Neuroscience, National Center of Neurology and Psychiatry,
Kodaira, Tokyo, Japan.
Address correspondence to Dr Hoffman, Research Center for Genetic
Medicine, Children’s National Medical Center, Department of
Integrative Systems Biology, George Washington University School
of Medicine, 111 Michigan Avenue NW, Washington, DC 20010.
being approximately average or within certain limits
Results: Weekly or biweekly systemic intravenous injections with a three-morpholino cocktail over the course of 5 to 22 weeks
induced therapeutic levels of dystrophin expression throughout the body, with an average of about 26% normal levels.
a substance that is used as a medicine or narcotic
Methods: We tested a series of antisense drugs singly and as cocktails, both in primary cell culture, and two in vivo delivery
methods (intramuscular injection and systemic intravenous injection).
the basic structural and functional unit of all organisms
Methods: We tested a series of antisense drugs singly and as cocktails, both in primary cell culture, and two in vivo delivery
methods (intramuscular injection and systemic intravenous injection).
relation with respect to comparative quantity or magnitude
Use of cocktails of morpholino, as
shown here, allows broader application of this approach to a greater proportion of DMD patients (90%) and also offers the
prospect of selecting deletions that optimize the functionality of the dystrophin protein.
an intermediate scale value regarded as normal or usual
Results: Weekly or biweekly systemic intravenous injections with a three-morpholino cocktail over the course of 5 to 22 weeks
induced therapeutic levels of dystrophin expression throughout the body, with an average of about 26% normal levels.
similar things placed in order or one after another
Methods: We tested a series of antisense drugs singly and as cocktails, both in primary cell culture, and two in vivo delivery
methods (intramuscular injection and systemic intravenous injection).