title: Ex vivo correction of selenoprotein N deficiency in rigid spine muscular dystrophy caused by a mutation in the selenocysteine codon.
abstract: Premature termination of translation due to nonsense mutations is a frequent cause of inherited diseases. Therefore, many efforts were invested in the development of strategies or compounds to selectively suppress this default. Selenoproteins are interesting candidates considering the idiosyncrasy of the amino acid selenocysteine (Sec) insertion mechanism. Here, we focused our studies on SEPN1, a selenoprotein gene whose mutations entail genetic disorders resulting in different forms of muscular diseases. Selective correction of a nonsense mutation at the Sec codon (UGA to UAA) was undertaken with a corrector tRNA(Sec) that was engineered to harbor a compensatory mutation in the anticodon. We demonstrated that its expression restored synthesis of a full-length selenoprotein N both in HeLa cells and in skin fibroblasts from a patient carrying the mutated Sec codon. Readthrough of the UAA codon was effectively dependent on the Sec insertion machinery, therefore being highly selective for this gene and unlikely to generate off-target effects. In addition, we observed that expression of the corrector tRNA(Sec) stabilized the mutated SEPN1 transcript that was otherwise more subject to degradation. In conclusion, our data provide interesting evidence that premature termination of translation due to nonsense mutations is amenable to correction, in the context of the specialized selenoprotein synthesis mechanism.
authors.affiliation: Architecture et Réactivité de l'ARN, Université Louis Pasteur de Strasbourg, CNRS, 67084 Strasbourg, France.
authors.forename: M
authors.initials: M
authors.lastname: Rederstorff
authors.forename: V
authors.initials: V
authors.lastname: Allamand
authors.forename: P
authors.initials: P
authors.lastname: Guicheney
authors.forename: C
authors.initials: C
authors.lastname: Gartioux
authors.forename: P
authors.initials: P
authors.lastname: Richard
authors.forename: D
authors.initials: D
authors.lastname: Chaigne
authors.forename: A
authors.initials: A
authors.lastname: Krol
authors.forename: A
authors.initials: A
authors.lastname: Lescure
mesh_terms: D003062:Codon / Q000737:chemistry; D018389:Codon, Nonsense; D005347:Fibroblasts / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D009124:Muscle Proteins / Q000096:biosynthesis / Q000172:deficiency* / Q000235:genetics*; D009134:Muscular Atrophy, Spinal / Q000235:genetics* / Q000378:metabolism; D012345:RNA, Transfer, Amino Acid-Specific / Q000235:genetics*; D017279:Selenocysteine / Q000378:metabolism; D051140:Selenoproteins / Q000096:biosynthesis / Q000172:deficiency* / Q000235:genetics*; D019076:Transgenes
chemical_list: D003062:Codon; D018389:Codon, Nonsense; D009124:Muscle Proteins; D012345:RNA, Transfer, Amino Acid-Specific; C435489:SELENON protein, human; D051140:Selenoproteins; C036928:tRNA, selenocysteine-; D017279:Selenocysteine