Abstract
Since its discovery in 1977, much has been known about RNA splicing and how it plays a central role in human development, function, and, notably, disease. Defects in RNA splicing account for at least 10% of all genetic disorders, with the number expected to increase as more information is uncovered on the contribution of noncoding genomic regions to disease. Splice modulation through the use of antisense oligonucleotides (AOs) has emerged as a promising avenue for the treatment of these disorders. In fact, two splice-switching AOs have recently obtained approval from the US Food and Drug Administration: eteplirsen (Exondys 51) for Duchenne muscular dystrophy, and nusinersen (Spinraza) for spinal muscular atrophy. These work by exon skipping and exon inclusion, respectively. In this chapter, we discuss the early development of AO-based splice modulation therapy—its invention, first applications, and its evolution into the approach we are now familiar with. We give a more extensive history of exon skipping in particular, as it is the splice modulation approach given the most focus in this book.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Chow LT, Gelinas RE, Broker TR, Roberts RJ (1977) An amazing sequence arrangement at the 5′ ends of adenovirus 2 messenger RNA. Cell 12:1–8
Berget SM, Moore C, Sharp PA (1977) Spliced segments at the 5′ terminus of adenovirus 2 late mRNA. Proc Natl Acad Sci U S A 74:3171–3175. https://doi.org/10.1073/pnas.74.8.3171
ENCODE Project Consortium (2012) An integrated encyclopedia of DNA elements in the human genome. Nature 489:57–74. https://doi.org/10.1038/nature11247
Keren H, Lev-Maor G, Ast G (2010) Alternative splicing and evolution: diversification, exon definition and function. Nat Rev Genet 11:345–355. https://doi.org/10.1038/nrg2776
Wang ET, Sandberg R, Luo S et al (2008) Alternative isoform regulation in human tissue transcriptomes. Nature 456:470–476. https://doi.org/10.1038/nature07509
Pan Q, Shai O, Lee LJ et al (2008) Deep surveying of alternative splicing complexity in the human transcriptome by high-throughput sequencing. Nat Genet 40:1413–1415. https://doi.org/10.1038/ng.259
Krawczak M, Thomas NST, Hundrieser B et al (2007) Single base-pair substitutions in exon-intron junctions of human genes: nature, distribution, and consequences for mRNA splicing. Hum Mutat 28:150–158. https://doi.org/10.1002/humu.20400
Douglas AGL, Wood MJA (2011) RNA splicing: disease and therapy. Brief Funct Genomics 10:151–164. https://doi.org/10.1093/bfgp/elr020
Hindorff LA, Sethupathy P, Junkins HA et al (2009) Potential etiologic and functional implications of genome-wide association loci for human diseases and traits. Proc Natl Acad Sci U S A 106:9362–9367. https://doi.org/10.1073/pnas.0903103106
López-Bigas N, Audit B, Ouzounis C et al (2005) Are splicing mutations the most frequent cause of hereditary disease? FEBS Lett 579:1900–1903. https://doi.org/10.1016/j.febslet.2005.02.047
Sazani P, Graziewicz M, Kole R (2007) Splice switching oligonucleotides as potential therapeutics. In: Antisense drug technol. CRC Press, Boca Raton, pp 89–114
Alberts B, Johnson A, Lewis J et al (2008) Molecular biology of the cell. In: Garland science, 5th edn. Taylor & Francis Group, New York
Chen M, Manley JL (2009) Mechanisms of alternative splicing regulation: insights from molecular and genomics approaches. Nat Rev Mol Cell Biol 10:741–754. https://doi.org/10.1038/nrm2777
Scotti MM, Swanson MS (2016) RNA mis-splicing in disease. Nat Rev Genet 17:19–32. https://doi.org/10.1038/nrg.2015.3
Turunen JJ, Niemelä EH, Verma B, Frilander MJ (2013) The significant other: splicing by the minor spliceosome. Wiley Interdiscip Rev RNA 4:61–76. https://doi.org/10.1002/wrna.1141
Corvelo A, Hallegger M, Smith CWJ, Eyras E (2010) Genome-wide association between branch point properties and alternative splicing. PLoS Comput Biol 6:e1001016. https://doi.org/10.1371/journal.pcbi.1001016
Schellenberg MJ, Ritchie DB, MacMillan AM (2008) Pre-mRNA splicing: a complex picture in higher definition. Trends Biochem Sci 33:243–246. https://doi.org/10.1016/j.tibs.2008.04.004
Coolidge CJ, Seely RJ, Patton JG (1997) Functional analysis of the polypyrimidine tract in pre-mRNA splicing. Nucleic Acids Res 25:888–896. https://doi.org/10.1093/nar/25.4.888
Stephenson ML, Zamecnik PC (1978) Inhibition of Rous sarcoma viral RNA translation by a specific oligodeoxyribonucleotide. Proc Natl Acad Sci U S A 75:285–288. https://doi.org/10.1073/pnas.75.1.285
Zamecnik PC, Stephenson ML (1978) Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide. Proc Natl Acad Sci U S A 75:280–284. https://doi.org/10.1073/pnas.75.1.280
Donis-Keller H (1979) Site specific enzymatic cleavage of RNA. Nucleic Acids Res 7:179–192. https://doi.org/10.1093/nar/7.1.179
Munroe SH (1988) Antisense RNA inhibits splicing of pre-mRNA in vitro. EMBO J 7:2523–2532
Mayeda A, Hayase Y, Inoue H et al (1990) Surveying cis-acting sequences of pre-mRNA by adding antisense 2’-O-methyl oligoribonucleotides to a splicing reaction. J Biochem 108:399–405
Bobst AM, Cerutti PA, Rottman F (1969) Structure of poly(2’-O-methyladenylic acid) at acidic and neutral pH. J Am Chem Soc 91:1246–1248. https://doi.org/10.1021/ja01033a054
Inoue H, Hayase Y, Imura A et al (1987) Synthesis and hybridization studies on two complementary nona(2’-O-methyl)ribonucleotides. Nucleic Acids Res 15:6131–6148. https://doi.org/10.1093/nar/15.15.6131
Inoue H, Hayase Y, Iwai S, Ohtsuka E (1987) Sequence-dependent hydrolysis of RNA using modified oligonucleotide splints and RNase H. Nucleic Acids Symp Ser 215:221–224. https://doi.org/10.1016/0014-5793(87)80171-0
Dominski Z, Kole R (1993) Restoration of correct splicing in thalassemic pre-mRNA by antisense oligonucleotides. Proc Natl Acad Sci U S A 90:8673–8677. https://doi.org/10.1021/ar00057a002
Thein SL (2013) The molecular basis of b-thalassemia. Cold Spring Harb Perspect Med 3:a011700–a011700. https://doi.org/10.1101/cshperspect.a011700
Eckstein F (1966) Nucleoside phosphorothioates. J Am Chem Soc 88:4292–4294. https://doi.org/10.1021/ja00970a054
Eckstein F (2000) Phosphorothioate oligodeoxynucleotides: what is their origin and what is unique about them? Antisense Nucleic Acid Drug Dev 10:117–121. https://doi.org/10.1089/oli.1.2000.10.117
Dowdy SF (2017) Overcoming cellular barriers for RNA therapeutics. Nat Biotechnol 35:222–229. https://doi.org/10.1038/nbt.3802
Miller PS, Yano J, Yano E et al (1979) Nonionic nucleic acid analogues. Synthesis and characterization of dideoxyribonucleoside methylphosphonates. Biochemistry 18:5134–5143
Sierakowska H, Sambade MJ, Agrawal S, Kole R (1996) Repair of thalassemic human beta-globin mRNA in mammalian cells by antisense oligonucleotides. Proc Natl Acad Sci U S A 93:12840–12844. https://doi.org/10.1073/pnas.93.23.12840
Sierakowska H, Montague M, Agrawal S, Kole R (1997) Restoration of ß-globin gene expression in mammalian cells by antisense oligonucleotides that modify the aberrant splicing patierns of thalassemic pre-mRNAs. Nucleosides and Nucleotides 16:1173–1182. https://doi.org/10.1080/07328319708006154
Friedman KJ, Kole J, Cohn JA et al (1999) Correction of aberrant splicing of the cystic fibrosis transmembrane conductance regulator ( CFTR ) gene by antisense oligonucleotides. J Biol Chem 274:36193–36199. https://doi.org/10.1074/jbc.274.51.36193
Scaffidi P, Misteli T (2005) Reversal of the cellular phenotype in the premature aging disease Hutchinson-Gilford progeria syndrome. Nat Med 11:440–445. https://doi.org/10.1038/nm1204
Vetrini F, Tammaro R, Bondanza S et al (2006) Aberrant splicing in the ocular albinism type 1 gene (OA1/GPR143) is corrected in vitro by morpholino antisense oligonucleotides. Hum Mutat 27:420–426. https://doi.org/10.1002/humu.20303
U.S. Food and Drug Administration (2016) FDA grants accelerated approval to first drug for Duchenne muscular dystrophy. http://www.fda.gov/NewsEvents/Newsroom/PressAnnouncements/ucm521263.htm. Accessed 30 Aug 2017
Emery AEH (1991) Population frequencies of inherited neuromuscular diseases-a world survey. Neuromuscul Disord 1:19–29
Mendell JR, Shilling C, Leslie ND et al (2012) Evidence-based path to newborn screening for Duchenne muscular dystrophy. Ann Neurol 71:304–313. https://doi.org/10.1002/ana.23528
Manzur A, Kinali M, Muntoni F (2008) Update on the management of Duchenne muscular dystrophy. Arch Dis Child 93:986–990. https://doi.org/10.1136/adc.2007.118141
Hoffman EP, Brown RH, Kunkel LM (1987) Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell 51:919–928. https://doi.org/10.1016/0092-8674(87)90579-4
Petrof BJ, Shrager JB, Stedman HH et al (1993) Dystrophin protects the sarcolemma from stresses developed during muscle contraction. Proc Natl Acad Sci U S A 90:3710–3714. https://doi.org/10.1073/pnas.90.8.3710
Koenig M, Hoffman EP, Bertelson CJ et al (1987) Complete cloning of the Duchenne muscular dystrophy (DMD) cDNA and preliminary genomic organization of the DMD gene in normal and affected individuals. Cell 50:509–517. https://doi.org/10.1016/0092-8674(87)90504-6
Roberts RG, Coffey AJ, Bobrow M, Bentley DR (1993) Exon structure of the human dystrophin gene. Genomics 16:536–538. https://doi.org/10.1006/geno.1993.1225
Monaco AP, Bertelson CJ, Liechti-Gallati S et al (1988) An explanation for the phenotypic differences between patients bearing partial deletions of the DMD locus. Genomics 2:90–95. https://doi.org/10.1016/0888-7543(88)90113-9
Koenig M, Beggs AH, Moyer M et al (1989) The molecular basis for Duchenne versus Becker muscular dystrophy: correlation of severity with type of deletion. Am J Hum Genet 45:498–506 doi: 10.1016/1
Matsuo M, Masumura T, Nakajima T et al (1990) A very small frame-shifting deletion within exon 19 of the Duchenne muscular dystrophy gene. Biochem Biophys Res Commun 170:963–967
Matsuo M, Masumura T, Nishio H et al (1991) Exon skipping during splicing of dystrophin mRNA precursor due to an intraexon deletion in the dystrophin gene of Duchenne muscular dystrophy Kobe. J Clin Invest 87:2127–2131. https://doi.org/10.1172/JCI115244
Takeshima Y, Nishio H, Sakamoto H et al (1995) Modulation of in vitro splicing of the upstream intron by modifying an intra-exon sequence which is deleted from the dystrophin gene in dystrophin Kobe. J Clin Invest 95:515–520. https://doi.org/10.1172/JCI117693
Nicholson LVB, Davison K, Johnson MA et al (1989) Dystrophin in skeletal muscle II. Immunoreactivity in patients with Xp21 muscular dystrophy. J Neurol Sci 94:137–146. https://doi.org/10.1016/0022-510X(89)90224-4
Hoffman EP, Morgan JE, Watkins SC, Partridge TA (1990) Somatic reversion/suppression of the mouse mdx phenotype in vivo. J Neurol Sci 99:9–25. https://doi.org/10.1016/0022-510X(90)90195-S
Klein CJ, Coovert DD, Bulman DE et al (1992) Somatic reversion/suppression in Duchenne muscular dystrophy (DMD): evidence supporting a frame-restoring mechanism in rare dystrophin-positive fibers. Am J Hum Genet 50:950–959
Thanh LT, Nguyen TM, Helliwell TR, Morris GE (1995) Characterization of revertant muscle fibers in Duchenne muscular dystrophy, using exon-specific monoclonal antibodies against dystrophin. Am J Hum Genet 56:725–731
Echigoya Y, Lee J, Rodrigues M et al (2013) Mutation types and aging differently affect revertant fiber expansion in dystrophic mdx and Mdx52 mice. PLoS One 8:e69194. https://doi.org/10.1371/journal.pone.0069194
Rodrigues M, Echigoya Y, Maruyama R et al (2016) Impaired regenerative capacity and lower revertant fibre expansion in dystrophin-deficient mdx muscles on DBA/2 background. Sci Rep 6:38371. https://doi.org/10.1038/srep38371
Pramono ZA, Takeshima Y, Alimsardjono H et al (1996) Induction of exon skipping of the dystrophin transcript in lymphoblastoid cells by transfecting an antisense oligodeoxynucleotide complementary to an exon recognition sequence. Biochem Biophys Res Commun 226:445–449
Sicinski P, Geng Y, Ryder-Cook AS et al (1989) The molecular basis of muscular dystrophy in the mdx mouse: a point mutation. Science 244:1578–1580
Dunckley MG, Manoharan M, Villiet P et al (1998) Modification of splicing in the dystrophin gene in cultured mdx muscle cells by antisense oligoribonucleotides. Hum Mol Genet 7:1083–1090
Wilton SD, Lloyd F, Carville K et al (1999) Specific removal of the nonsense mutation from the mdx dystrophin mRNA using antisense oligonucleotides. Neuromuscul Disord 9:330–338. https://doi.org/10.1016/S0960-8966(99)00010-3
van Deutekom JC, Bremmer-Bout M, Janson AA et al (2001) Antisense-induced exon skipping restores dystrophin expression in DMD patient derived muscle cells. Hum Mol Genet 10:1547–1554. https://doi.org/10.1093/HMG/10.15.1547
Takeshima Y, Wada H, Yagi M et al (2001) Oligonucleotides against a splicing enhancer sequence led to dystrophin production in muscle cells from a Duchenne muscular dystrophy patient. Brain and Development 23:788–790. https://doi.org/10.1016/S0387-7604(01)00326-6
Aartsma-Rus A, Janson AAM, Kaman WE et al (2003) Therapeutic antisense-induced exon skipping in cultured muscle cells from six different DMD patients. Hum Mol Genet 12:907–914
Aartsma-Rus A, Janson AA, Kaman WE et al (2004) Antisense-induced multiexon skipping for Duchenne muscular dystrophy makes more sense. Am J Hum Genet 74:83–92. https://doi.org/10.1086/381039
Aartsma-Rus A, Kaman WE, Bremmer-Bout M et al (2004) Comparative analysis of antisense oligonucleotide analogs for targeted DMD exon 46 skipping in muscle cells. Gene Ther 11:1391–1398. https://doi.org/10.1038/sj.gt.3302313
Surono A, Van Khanh T, Takeshima Y et al (2004) Chimeric RNA/ethylene-bridged nucleic acids promote dystrophin expression in myocytes of duchenne muscular dystrophy by inducing skipping of the nonsense mutation-encoding exon. Hum Gene Ther 15:749–757. https://doi.org/10.1089/1043034041648444
Aartsma-Rus A, Janson AAM, van Ommen G-JB, van Deutekom JCT (2007) Antisense-induced exon skipping for duplications in Duchenne muscular dystrophy. BMC Med Genet 8:43. https://doi.org/10.1186/1471-2350-8-43
Mann CJ, Honeyman K, Cheng AJ et al (2001) Antisense-induced exon skipping and synthesis of dystrophin in the mdx mouse. Proc Natl Acad Sci U S A 98:42–47. https://doi.org/10.1073/pnas.011408598
Lu QL, Mann CJ, Lou F et al (2003) Functional amounts of dystrophin produced by skipping the mutated exon in the mdx dystrophic mouse. Nat Med 9:1009–1014. https://doi.org/10.1038/nm897
Summerton JE (2017) Invention and early history of morpholinos: from pipe dream to practical products. Methods Mol Biol 1565:1–15. https://doi.org/10.1007/978-1-4939-6817-6_1
Summerton J, Weller D (1997) Morpholino antisense oligomers: design, preparation, and properties. Antisense Nucleic Acid Drug Dev 7:187–195. https://doi.org/10.1089/oli.1.1997.7.187
Lee JJA, Yokota T (2013) Antisense therapy in neurology. J Pers Med 3:144–176. https://doi.org/10.3390/jpm3030144
Moulton JD (2016) Guide for morpholino users: toward therapeutics. J Drug Discov Dev Deliv 3:1023
Gebski BL, Mann CJ, Fletcher S, Wilton SD (2003) Morpholino antisense oligonucleotide induced dystrophin exon 23 skipping in mdx mouse muscle. Hum Mol Genet 12:1801–1811. https://doi.org/10.1093/hmg/ddg196
Schmajuk G, Sierakowska H, Kole R (1999) Antisense oligonucleotides with different backbones. Modification of splicing pathways and efficacy of uptake. J Biol Chem 274:21783–21789
Lacerra G, Sierakowska H, Carestia C et al (2000) Restoration of hemoglobin a synthesis in erythroid cells from peripheral blood of thalassemic patients. Proc Natl Acad Sci 97:9591–9596. https://doi.org/10.1073/pnas.97.17.9591
Suwanmanee T, Sierakowska H, Lacerra G et al (2002) Restoration of human beta-globin gene expression in murine and human IVS2-654 thalassemic erythroid cells by free uptake of antisense oligonucleotides. Mol Pharmacol 62:545–553
Suwanmanee T, Sierakowska H, Fucharoen S, Kole R (2002) Repair of a splicing defect in erythroid cells from patients with beta-thalassemia/HbE disorder. Mol Ther 6:718–726
Wells KE, Fletcher S, Mann CJ et al (2003) Enhanced in vivo delivery of antisense oligonucleotides to restore dystrophin expression in adult mdx mouse muscle. FEBS Lett 552:145–149
Graham IR, Hill VJ, Manoharan M et al (2004) Towards a therapeutic inhibition of dystrophin exon 23 splicing in mdx mouse muscle induced by antisense oligoribonucleotides (splicomers): target sequence optimisation using oligonucleotide arrays. J Gene Med 6:1149–1158. https://doi.org/10.1002/jgm.603
Lu QL, Rabinowitz A, Chen YC et al (2005) Systemic delivery of antisense oligoribonucleotide restores dystrophin expression in body-wide skeletal muscles. Proc Natl Acad Sci U S A 102:198–203. https://doi.org/10.1073/pnas.0406700102
Alter J, Lou F, Rabinowitz A et al (2006) Systemic delivery of morpholino oligonucleotide restores dystrophin expression bodywide and improves dystrophic pathology. Nat Med 12:175–177. https://doi.org/10.1038/nm1345
Yu X, Bao B, Echigoya Y, Yokota T (2015) Dystrophin-deficient large animal models: translational research and exon skipping. Am J Transl Res 7:1314–1331
Shimatsu Y, Katagiri K, Furuta T et al (2003) Canine X-linked muscular dystrophy in Japan (CXMDJ). Exp Anim 52:93–97
Yokota T, Lu Q-L, Partridge T et al (2009) Efficacy of systemic morpholino exon-skipping in Duchenne dystrophy dogs. Ann Neurol 65:667–676. https://doi.org/10.1002/ana.21627
McClorey G, Moulton HM, Iversen PL et al (2006) Antisense oligonucleotide-induced exon skipping restores dystrophin expression in vitro in a canine model of DMD. Gene Ther 13:1373–1381. https://doi.org/10.1038/sj.gt.3302800
Takeshima Y, Yagi M, Wada H et al (2006) Intravenous infusion of an antisense oligonucleotide results in exon skipping in muscle dystrophin mRNA of Duchenne muscular dystrophy. Pediatr Res 59:690–694. https://doi.org/10.1203/01.pdr.0000215047.51278.7c
van Deutekom JC, Janson AA, Ginjaar IB et al (2007) Local dystrophin restoration with antisense oligonucleotide PRO051. N Engl J Med 357:2677–2686. https://doi.org/10.1056/NEJMoa073108
Kinali M, Arechavala-Gomeza V, Feng L et al (2009) Local restoration of dystrophin expression with the morpholino oligomer AVI-4658 in Duchenne muscular dystrophy: a single-blind, placebo-controlled, dose-escalation, proof-of-concept study. Lancet Neurol 8:918–928. https://doi.org/10.1016/S1474-4422(09)70211-X
Mendell JR, Rodino-Klapac LR, Sahenk Z et al (2013) Eteplirsen for the treatment of Duchenne muscular dystrophy. Ann Neurol 74:637–647. https://doi.org/10.1002/ana.23982
Kesselheim AS, Avorn J (2016) Approving a problematic muscular dystrophy drug: implications for FDA policy. JAMA 316(22):2357–2358. https://doi.org/10.1001/jama.2016.16437
Lim KRQ, Maruyama R, Yokota T (2017) Eteplirsen in the treatment of Duchenne muscular dystrophy. Drug Des Devel Ther 11:533–545. https://doi.org/10.2147/DDDT.S97635
U.S. Food and Drug Administration: Center for Drug Evaluation and Research (2016) Summary review, application number: 206488Orig1s000. http://www.accessdata.fda.gov/drugsatfda_docs/nda/2016/206488_summary review_Redacted.pdf. Accessed 30 Aug 2017
Echigoya Y, Lim KRQ, Trieu N et al (2017) Quantitative antisense screening and optimization for exon 51 skipping in duchenne muscular dystrophy. Mol Ther 25(11):2561–2572. https://doi.org/10.1016/j.ymthe.2017.07.014
Guncay A, Yokota T (2015) Antisense oligonucleotide drugs for Duchenne muscular dystrophy: how far have we come and what does the future hold? Future Med Chem 7:1631–1635. https://doi.org/10.4155/fmc.15.116
Nguyen Q, Yokota T (2017) Immortalized muscle cell model to test the exon skipping efficacy for duchenne muscular dystrophy. J Pers Med 7:13. https://doi.org/10.3390/jpm7040013
Taylor JK, Zhang QQ, Wyatt JR, Dean NM (1999) Induction of endogenous Bcl-xS through the control of Bcl-x pre-mRNA splicing by antisense oligonucleotides. Nat Biotechnol 17:1097–1100. https://doi.org/10.1038/15079
Martin P (1995) Ein neuer Zugang zu 2?-O-Alkylribonucleosiden und Eigenschaften deren Oligonucleotide. Helv Chim Acta 78:486–504. https://doi.org/10.1002/hlca.19950780219
Mercatante DR, Kole R (2002) Control of alternative splicing by antisense oligonucleotides as a potential chemotherapy: effects on gene expression. Biochim Biophys Acta 1587:126–132. https://doi.org/10.1016/S0925-4439(02)00075-3
Mercatante DR, Bortner CD, Cidlowski JA, Kole R (2001) Modification of alternative splicing of Bcl-x pre-mRNA in prostate and breast cancer cells. Analysis of apoptosis and cell death. J Biol Chem 276:16411–16417. https://doi.org/10.1074/jbc.M009256200
McGrath JA, Ashton GHS, Mellerio JE et al (1999) Moderation of phenotypic severity in dystrophic and junctional forms of epidermolysis bullosa through in-frame skipping of exons containing non-sense or frameshift mutations. J Invest Dermatol 113:314–321. https://doi.org/10.1046/j.1523-1747.1999.00709.x
Kalbfuss B, Mabon SA, Misteli T (2001) Correction of alternative splicing of tau in frontotemporal dementia and parkinsonism linked to chromosome 17. J Biol Chem 276:42986–42993. https://doi.org/10.1074/jbc.M105113200
Renshaw J, Orr RM, Walton MI et al (2004) Disruption of WT1 gene expression and exon 5 splicing following cytotoxic drug treatment: antisense down-regulation of exon 5 alters target gene expression and inhibits cell survival. Mol Cancer Ther 3:1467–1484
Meijboom KE, Wood MJA, McClorey G (2017) Splice-switching therapy for spinal muscular atrophy. Genes (Basel) 8(6):E161. https://doi.org/10.3390/genes8060161
Arnold WD, Kassar D, Kissel JT (2015) Spinal muscular atrophy: diagnosis and management in a new therapeutic era. Muscle Nerve 51:157–167. https://doi.org/10.1002/mus.24497
Wirth B, Herz M, Wetter A et al (1999) Quantitative analysis of survival motor neuron copies: identification of subtle SMN1 mutations in patients with spinal muscular atrophy, genotype-phenotype correlation, and implications for genetic counseling. Am J Hum Genet 64:1340–1356. https://doi.org/10.1086/302369
Mailman MD, Heinz JW, Papp AC et al (2002) Molecular analysis of spinal muscular atrophy and modification of the phenotype by SMN2. Genet Med 4:20–26. https://doi.org/10.1097/00125817-200201000-00004
Lorson CL, Hahnen E, Androphy EJ, Wirth B (1999) A single nucleotide in the SMN gene regulates splicing and is responsible for spinal muscular atrophy. Proc Natl Acad Sci U S A 96:6307–6311
Monani UR, Lorson CL, Parsons DW et al (1999) A single nucleotide difference that alters splicing patterns distinguishes the SMA gene SMN1 from the copy gene SMN2. Hum Mol Genet 8:1177–1183
Lim SR, Hertel KJ (2001) Modulation of survival motor neuron pre-mRNA splicing by inhibition of alternative 3′ splice site pairing. J Biol Chem 276:45476–45483. https://doi.org/10.1074/jbc.M107632200
Miyajima H, Miyaso H, Okumura M et al (2002) Identification of a cis-acting element for the regulation of SMN exon 7 splicing. J Biol Chem 277:23271–23277. https://doi.org/10.1074/jbc.M200851200
Skordis LA, Dunckley MG, Yue B et al (2003) Bifunctional antisense oligonucleotides provide a trans-acting splicing enhancer that stimulates SMN2 gene expression in patient fibroblasts. Proc Natl Acad Sci U S A 100:4114–4119. https://doi.org/10.1073/pnas.0633863100
Touznik A, Maruyama R, Hosoki K et al (2017) LNA/DNA mixmer-based antisense oligonucleotides correct alternative splicing of the SMN2 gene and restore SMN protein expression in type 1 SMA fibroblasts. Sci Rep 7:3672. https://doi.org/10.1038/s41598-017-03850-2
Singh NK, Singh NN, Androphy EJ, Singh RN (2006) Splicing of a critical exon of human survival motor neuron is regulated by a unique silencer element located in the last intron. Mol Cell Biol 26:1333–1346. https://doi.org/10.1128/MCB.26.4.1333-1346.2006
Hua Y, Vickers TA, Okunola HL et al (2008) Antisense masking of an hnRNP A1/A2 intronic splicing silencer corrects SMN2 splicing in transgenic mice. Am J Hum Genet 82:834–848. https://doi.org/10.1016/j.ajhg.2008.01.014
Hua Y, Sahashi K, Hung G et al (2010) Antisense correction of SMN2 splicing in the CNS rescues necrosis in a type III SMA mouse model. Genes Dev 24:1634–1644. https://doi.org/10.1101/gad.1941310
Passini MA, Bu J, Richards AM et al (2011) Antisense oligonucleotides delivered to the mouse CNS ameliorate symptoms of severe spinal muscular atrophy. Sci Transl Med 3:72ra18. https://doi.org/10.1126/scitranslmed.3001777
Chiriboga CA, Swoboda KJ, Darras BT et al (2016) Results from a phase 1 study of nusinersen (ISIS-SMN Rx ) in children with spinal muscular atrophy. Neurology 86:890–897. https://doi.org/10.1212/WNL.0000000000002445
U.S. Food and Drug Administration (2016) FDA approves first drug for spinal muscular atrophy. https://www.fda.gov/newsevents/newsroom/pressannouncements/ucm534611.htm. Accessed 29 Nov 2017
Juliano R, Bauman J, Kang H, Ming X (2009) Biological barriers to therapy with antisense and siRNA oligonucleotides. Mol Pharm 6:686–695. https://doi.org/10.1021/mp900093r
Khvorova A, Watts JK (2017) The chemical evolution of oligonucleotide therapies of clinical utility. Nat Biotechnol 35:238–248. https://doi.org/10.1038/nbt.3765
Nielsen PE, Egholm M, Berg RH, Buchardt O (1991) Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide. Science 254:1497–1500
Koshkin AA, Singh SK, Nielsen P et al (1998) LNA (locked nucleic acids): synthesis of the adenine, cytosine, guanine, 5-methylcytosine, thymine and uracil bicyclonucleoside monomers, oligomerisation, and unprecedented nucleic acid recognition. Tetrahedron 54:3607–3630. https://doi.org/10.1016/S0040-4020(98)00094-5
Obika S, Nanbu D, Hari Y et al (1998) Stability and structural features of the duplexes containing nucleoside analogues with a fixed N-type conformation, 2′-O,4′-C-methyleneribonucleosides. Tetrahedron Lett 39:5401–5404. https://doi.org/10.1016/S0040-4039(98)01084-3
Steffens R, Leumann CJ (1999) Synthesis and thermodynamic and biophysical properties of tricyclo-DNA. J Am Chem Soc 121:3249–3255. https://doi.org/10.1021/ja983570w
Seth PP, Siwkowski A, Allerson CR et al (2008) Design, synthesis and evaluation of constrained methoxyethyl (cMOE) and constrained ethyl (cEt) nucleoside analogs. Nucleic Acids Symp Ser (Oxf) 52:553–554. https://doi.org/10.1093/nass/nrn280
Moulton HM, Moulton JD (2010) Morpholinos and their peptide conjugates: therapeutic promise and challenge for Duchenne muscular dystrophy. Biochim Biophys Acta 1798:2296–2303. https://doi.org/10.1016/j.bbamem.2010.02.012
Moulton HM, Moulton JD (2003) Peptide-assisted delivery of steric-blocking antisense oligomers. Curr Opin Mol Ther 5:123–132
Echigoya Y, Nakamura A, Nagata T et al (2017) Effects of systemic multiexon skipping with peptide-conjugated morpholinos in the heart of a dog model of Duchenne muscular dystrophy. Proc Natl Acad Sci 114:4213–4218. https://doi.org/10.1073/pnas.1613203114
Echigoya Y, Mouly V, Garcia L et al (2015) In silico screening based on predictive algorithms as a design tool for exon skipping oligonucleotides in duchenne muscular dystrophy. PLoS One 10:e0120058. https://doi.org/10.1371/journal.pone.0120058
O’Leary DA, Sharif O, Anderson P et al (2009) Identification of small molecule and genetic modulators of AON-induced dystrophin exon skipping by high-throughput screening. PLoS One 4:e8348. https://doi.org/10.1371/journal.pone.0008348
Naryshkin N, Dakka A, Gabbeta V et al (2015) Small molecule compounds that promote exon skipping in the DMD gene. Neuromuscul Disord 25:S261. https://doi.org/10.1016/j.nmd.2015.06.275
Kendall GC, Mokhonova EI, Moran M et al (2012) Dantrolene enhances antisense-mediated exon skipping in human and mouse models of Duchenne muscular dystrophy. Sci Transl Med 4:164ra160. https://doi.org/10.1126/scitranslmed.3005054
Han G, Gu B, Cao L et al (2016) Hexose enhances oligonucleotide delivery and exon skipping in dystrophin-deficient mdx mice. Nat Commun 7:10981. https://doi.org/10.1038/ncomms10981
Yokota T, Takeda S, Lu Q-L et al (2009) A renaissance for antisense oligonucleotide drugs in neurology: exon skipping breaks new ground. Arch Neurol 66:32–38. https://doi.org/10.1001/archneurol.2008.540
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Lim, K.R.Q., Yokota, T. (2018). Invention and Early History of Exon Skipping and Splice Modulation. 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_1
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8651-4_1
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8650-7
Online ISBN: 978-1-4939-8651-4
eBook Packages: Springer Protocols