{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T13:40:17Z","timestamp":1791207617029,"version":"4.1.0"},"reference-count":36,"publisher":"National Academy of Sciences","issue":"7","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2003,4]]},"abstract":"<jats:p>\n                    The multiplicity of proteins compared with genes in mammals owes much to alternative splicing. Splicing signals are so subtle and complex that small perturbations may allow the production of new mRNA variants. However, the flexibility of splicing can also be a liability, and several genetic diseases result from single-base changes that cause exons to be skipped during splicing. Conventional oligonucleotide strategies can block reactions but cannot restore splicing. We describe here a method by which the use of a defective exon was restored. Spinal muscular atrophy (SMA) results from mutations of the\n                    <jats:italic>Survival Motor Neuron<\/jats:italic>\n                    (\n                    <jats:italic>SMN<\/jats:italic>\n                    ) gene. Mutations of\n                    <jats:italic>SMN1<\/jats:italic>\n                    cause SMA, whereas\n                    <jats:italic>SMN2<\/jats:italic>\n                    acts as a modifying gene. The two genes undergo alternative splicing with\n                    <jats:italic>SMN1<\/jats:italic>\n                    , producing an abundance of full-length mRNA transcripts, whereas\n                    <jats:italic>SMN2<\/jats:italic>\n                    predominantly produces exon 7-deleted transcripts. This discrepancy is because of a single nucleotide difference in\n                    <jats:italic>SMN2<\/jats:italic>\n                    exon 7, which disrupts an exonic splicing enhancer containing an SF2\/ASF binding site. We have designed oligoribonucleotides that are complementary to exon 7 and contain exonic splicing enhancer motifs to provide trans-acting enhancers. These tailed oligoribonucleotides increased\n                    <jats:italic>SMN2<\/jats:italic>\n                    exon 7 splicing\n                    <jats:italic>in vitro<\/jats:italic>\n                    and rescued the incorporation of\n                    <jats:italic>SMN2<\/jats:italic>\n                    exon 7 in SMA patient fibroblasts. This treatment also resulted in the partial restoration of gems, intranuclear structures containing SMN protein that are severely reduced in patients with SMA. The use of tailed antisense oligonucleotides to recruit positively acting factors to stimulate a splicing reaction may have therapeutic applications for genetic disorders, such as SMA, in which splicing patterns are altered.\n                  <\/jats:p>","DOI":"10.1073\/pnas.0633863100","type":"journal-article","created":{"date-parts":[[2003,4,1]],"date-time":"2003-04-01T14:25:08Z","timestamp":1049207108000},"page":"4114-4119","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":233,"title":["Bifunctional antisense oligonucleotides provide a trans-acting splicing enhancer that stimulates\n                    <i>SMN2<\/i>\n                    gene expression in patient fibroblasts"],"prefix":"10.1073","volume":"100","author":[{"given":"Leigh A.","family":"Skordis","sequence":"first","affiliation":[{"name":"Dubowitz Neuromuscular Centre, Imperial College London, Hammersmith Hospital, London W12 0NN, United Kingdom Europe; and Department of Biochemistry, University of Leicester, Leicester LE1 7RH, United Kingdom Europe"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew G.","family":"Dunckley","sequence":"additional","affiliation":[{"name":"Dubowitz Neuromuscular Centre, Imperial College London, Hammersmith Hospital, London W12 0NN, United Kingdom Europe; and Department of Biochemistry, University of Leicester, Leicester LE1 7RH, United Kingdom Europe"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baigong","family":"Yue","sequence":"additional","affiliation":[{"name":"Dubowitz Neuromuscular Centre, Imperial College London, Hammersmith Hospital, London W12 0NN, United Kingdom Europe; and Department of Biochemistry, University of Leicester, Leicester LE1 7RH, United Kingdom Europe"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian C.","family":"Eperon","sequence":"additional","affiliation":[{"name":"Dubowitz Neuromuscular Centre, Imperial College London, Hammersmith Hospital, London W12 0NN, United Kingdom Europe; and Department of Biochemistry, University of Leicester, Leicester LE1 7RH, United Kingdom Europe"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"Muntoni","sequence":"additional","affiliation":[{"name":"Dubowitz Neuromuscular Centre, Imperial College London, Hammersmith Hospital, London W12 0NN, United Kingdom Europe; and Department of Biochemistry, University of Leicester, Leicester LE1 7RH, United Kingdom Europe"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2003,3,17]]},"reference":[{"key":"e_1_3_3_1_2","volume-title":"Muscle Disorders in Childhood","author":"Dubowitz V","year":"1995","unstructured":"V Dubowitz Muscle Disorders in Childhood (Saunders, 2nd Ed., London, 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