title: Redirecting splicing with bifunctional oligonucleotides.
abstract: Ectopic modulators of alternative splicing are important tools to study the function of splice variants and for correcting mis-splicing events that cause human diseases. Such modulators can be bifunctional oligonucleotides made of an antisense portion that determines target specificity, and a non-hybridizing tail that recruits proteins or RNA/protein complexes that affect splice site selection (TOSS and TOES, respectively, for targeted oligonucleotide silencer of splicing and targeted oligonucleotide enhancer of splicing). The use of TOSS and TOES has been restricted to a handful of targets. To generalize the applicability and demonstrate the robustness of TOSS, we have tested this approach on more than 50 alternative splicing events. Moreover, we have developed an algorithm that can design active TOSS with a success rate of 80%. To produce bifunctional oligonucleotides capable of stimulating splicing, we built on the observation that binding sites for TDP-43 can stimulate splicing and improve U1 snRNP binding when inserted downstream from 5' splice sites. A TOES designed to recruit TDP-43 improved exon 7 inclusion in SMN2. Overall, our study shows that bifunctional oligonucleotides can redirect splicing on a variety of genes, justifying their inclusion in the molecular arsenal that aims to alter the production of splice variants.
authors.affiliation: Laboratory of Functional Genomics and Research Centre on RNA Biology of the Université de Sherbrooke, Sherbrooke, Quebec J1E 4K8, Canada, Department of Biochemistry, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Quebec J1E 4K8, Canada and Department of Microbiology and Infectious Diseases, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Quebec J1E 4K8, Canada.
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Brosseau
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Lucier
authors.forename: Andrée-Anne
authors.initials: AA
authors.lastname: Lamarche
authors.forename: Lulzim
authors.initials: L
authors.lastname: Shkreta
authors.forename: Daniel
authors.initials: D
authors.lastname: Gendron
authors.forename: Elvy
authors.initials: E
authors.lastname: Lapointe
authors.forename: Philippe
authors.initials: P
authors.lastname: Thibault
authors.forename: Eric
authors.initials: E
authors.lastname: Paquet
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Perreault
authors.forename: Sherif
authors.initials: S
authors.lastname: Abou Elela
authors.forename: Benoit
authors.initials: B
authors.lastname: Chabot
mesh_terms: D000465:Algorithms; D017398:Alternative Splicing; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism; D005091:Exons; D006367:HeLa Cells; D006801:Humans; D009841:Oligonucleotides / Q000737:chemistry*; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D022821:RNA Splice Sites; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D022821:RNA Splice Sites; D017412:Ribonucleoprotein, U1 Small Nuclear