title: Efficient in vivo gene expression by trans-splicing adeno-associated viral vectors.
abstract: Although adeno-associated virus (AAV)-mediated gene therapy has been hindered by the small viral packaging capacity of the vector, trans-splicing AAV vectors are able to package twice the size of the vector genome. Unfortunately, the efficiency of current trans-splicing vectors is very low. Here we show that rational design of the gene splitting site has a profound influence on trans-splicing vector-mediated gene expression. Using mRNA accumulation as a guide, we generated a set of efficient trans-splicing vectors and achieved widespread expression of the 6-kb DeltaH2-R19 mini-dystrophin gene in skeletal muscle of mdx mice, a model for Duchenne muscular dystrophy. The dystrophic phenotype was ameliorated in both adult and aged mice. This demonstrates the use of trans-splicing vectors to efficiently express a large therapeutic structural protein. This strategy should be applicable to other large therapeutic genes or large transcription regulatory elements.
authors.affiliation: Department of Molecular Microbiology and Immunology, University of Missouri School of Medicine, 1 Hospital Dr., Room M610G, MSB, Columbia, Missouri 65212, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Lai
authors.forename: Yongping
authors.initials: Y
authors.lastname: Yue
authors.forename: Mingju
authors.initials: M
authors.lastname: Liu
authors.forename: Arkasubhra
authors.initials: A
authors.lastname: Ghosh
authors.forename: John F
authors.initials: JF
authors.lastname: Engelhardt
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Chamberlain
authors.forename: Dongsheng
authors.initials: D
authors.lastname: Duan
mesh_terms: D000818:Animals; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D016189:Dystrophin / Q000235:genetics; D005091:Exons; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D007438:Introns; D008297:Male; D051379:Mice; D018101:Mice, Inbred mdx; D015233:Models, Statistical; D018482:Muscle, Skeletal / Q000378:metabolism; D009132:Muscles / Q000473:pathology; D010641:Phenotype; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism; D020040:Trans-Splicing; D014158:Transcription, Genetic
chemical_list: D016189:Dystrophin; D012333:RNA, Messenger