title: A novel therapeutic approach for genetic diseases by introduction of suppressor tRNA.
abstract: The appearance of the premature translation termination codons (PTCs) in the transcript is the major cause of human genetic diseases. PTC-containing transcripts are rapidly degraded through nonsense-mediated decay (NMD) pathway. If such mRNA transcripts were translated in frame like normal transcripts, it would afford not only restoration of the level of full-length protein but also prevention of mRNA degradation by the NMD pathway. Here we describe a novel approach to read through PTC-containing mRNAs using suppressor tRNA that is introduced to cells by transfection. Luciferase reporter gene assay showed that nonsense and four-base codons were suppressed by the corresponding suppressor tRNAs derived from human tRNA(Ser). We also demonstrated that transfection of the suppressor tRNA to Ullrich disease fibroblasts, possessing a frameshift mutation in the collagen VI alpha2 gene, induced the upregulation of the collagen VI alpha2 mRNA and accumulation of the collagen VI protein. PTC suppression potentially provides a novel therapeutic means to rescue various PTC-related diseases.
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Tokyo 153-8904, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Sako
authors.forename: Fusako
authors.initials: F
authors.lastname: Usuki
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Suga
mesh_terms: D000926:Anticodon / Q000737:chemistry; D018389:Codon, Nonsense; D024142:Collagen Type VI / Q000096:biosynthesis / Q000235:genetics*; D016368:Frameshift Mutation; D017930:Genes, Reporter; D006801:Humans; D009136:Muscular Dystrophies / Q000235:genetics / Q000628:therapy; D010443:Peptide Chain Termination, Translational; D012333:RNA, Messenger / Q000096:biosynthesis; D012343:RNA, Transfer / Q000737:chemistry*; D012362:RNA, Transfer, Ser / Q000737:chemistry; D013489:Suppression, Genetic
chemical_list: D000926:Anticodon; D018389:Codon, Nonsense; D024142:Collagen Type VI; D012333:RNA, Messenger; D012362:RNA, Transfer, Ser; D012343:RNA, Transfer