title: Cis-splicing and translation of the pre-trans-splicing molecule combine with efficiency in spliceosome-mediated RNA trans-splicing.
abstract: Muscular dystrophies are a group of genetically distinct diseases for which no treatment exists. While gene transfer approach is being tested for several of these diseases, such strategies can be hampered when the size of the corresponding complementary DNA (cDNA) exceeds the packaging capacity of adeno-associated virus vectors. This issue concerns, in particular, dysferlinopathies and titinopathies that are due to mutations in the dysferlin (DYSF) and titin (TTN) genes. We investigated the efficacy of RNA trans-splicing as a mode of RNA therapy for these two types of diseases. Results obtained with RNA trans-splicing molecules designed to target the 3' end of mouse titin and human dysferlin pre-mRNA transcripts indicated that trans-splicing of pre-mRNA generated from minigene constructs or from the endogenous genes was achieved. Collectively, these results provide the first demonstration of DYSF and TTN trans-splicing reprogramming in vitro and in vivo. However, in addition to these positive results, we uncovered a possible issue of the technique in the form of undesirable translation of RNA pre-trans-splicing molecules, directly from open reading frames present on the molecule or associated with internal alternative cis-splicing. These events may hamper the efficiency of the trans-splicing process and/or lead to toxicity.
authors.affiliation: Généthon, CNRS UMR8587, 1, rue de l'Internationale, Evry, France.
authors.forename: François
authors.initials: F
authors.lastname: Monjaret
authors.affiliation: Généthon, CNRS UMR8587, 1, rue de l'Internationale, Evry, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Bourg
authors.affiliation: Généthon, CNRS UMR8587, 1, rue de l'Internationale, Evry, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Suel
authors.affiliation: Généthon, CNRS UMR8587, 1, rue de l'Internationale, Evry, France.
authors.forename: Carinne
authors.initials: C
authors.lastname: Roudaut
authors.affiliation: Généthon, CNRS UMR8587, 1, rue de l'Internationale, Evry, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Le Roy
authors.affiliation: Généthon, CNRS UMR8587, 1, rue de l'Internationale, Evry, France. Electronic address: richard@genethon.fr.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Richard
authors.affiliation: Généthon, CNRS UMR8587, 1, rue de l'Internationale, Evry, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Charton
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D002460:Cell Line; D000073939:Dysferlin; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D058990:Molecular Targeted Therapy; D009136:Muscular Dystrophies / Q000235:genetics / Q000628:therapy*; D016366:Open Reading Frames; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism*; D012322:RNA Precursors / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism*; D020040:Trans-Splicing
chemical_list: C497505:Dysf protein, mouse; D000073939:Dysferlin; D008565:Membrane Proteins; D012322:RNA Precursors; D012333:RNA, Messenger; D011494:Protein Kinases; C579730:titin protein, mouse