title: Design of trans-splicing adeno-associated viral vectors for Duchenne muscular dystrophy gene therapy.
abstract: The development of trans-splicing vectors opens the door for delivering a large therapeutic gene with adeno-associated viral vectors (AAV). One potential application is to deliver the 6 kb mini-dystrophin gene for Duchenne muscular dystrophy (DMD) gene therapy. However, early attempts have been very disappointing because of low transduction efficiency. We have recently identified mRNA accumulation as a critical barrier for the trans-splicing AAV vectors. This barrier can be overcome by rational selection of the gene splitting site. Here we outline a detailed RNase protection assay-based strategy to determine the optimal gene splitting site for the mini-dystrophin gene. We also provide methods to evaluate transduction efficiency of the mini-dystrophin trans-splicing vectors in mdx mouse, a model for DMD.
authors.affiliation: Department of Molecular Microbiology and Immunology, University of Missouri, Columbia, MO, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Lai
authors.forename: Dejia
authors.initials: D
authors.lastname: Li
authors.forename: Yongping
authors.initials: Y
authors.lastname: Yue
authors.forename: Dongsheng
authors.initials: D
authors.lastname: Duan
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics*; D016189:Dystrophin / Q000737:chemistry / Q000235:genetics / Q000627:therapeutic use; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D051379:Mice; D018101:Mice, Inbred mdx; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000628:therapy*; D011323:Primates / Q000235:genetics; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics; D020040:Trans-Splicing / Q000235:genetics*; D014161:Transduction, Genetic
chemical_list: D016189:Dystrophin; D020033:Protein Isoforms