development of transgenic mice containing an introduced stop codon on the human methylmalonyl-coa mutase locus发展转基因老鼠包含一个终止密码子介绍人类methylmalonyl-coa变位酶位点.pdfVIP

Development of Transgenic Mice Containing an Introduced Stop Codon on the Human Methylmalonyl- CoA Mutase Locus 1 1 1,2 1 1 ¤ Nicole E. Buck , Harriet Dashnow , James J. Pitt , Leonie R. Wood , Heidi L. Peters * 1 Metabolic Research, Murdoch Childrens Research Institute, The University of Melbourne Department of Paediatrics, Parkville, Victoria, Australia, 2 VCGS Pathology, Murdoch Childrens Research Institute, Parkville, Victoria, Australia Abstract The mutation R403stop was found in an individual with mut0 methylmalonic aciduria (MMA) which resulted from a single base change of CRT in exon 6 of the methylmalonyl-CoA mutase gene (producing a TGA stop codon). In order to accurately model the human MMA disorder we introduced this mutation onto the human methylmalonyl-CoA mutase locus of a bacterial artificial chromosome. A mouse model was developed using this construct. The transgene was found to be intact in the mouse model, with 7 copies integrated at a single site in chromosome 3. The phenotype of the hemizygous mouse was unchanged until crossed against a methylmalonyl-CoA mutase knockout mouse. Pups with no endogenous mouse methylmalonyl-CoA mutase and one copy of the transgene became ill and died within 24 hours. This severe phenotype could be partially rescued by the addition of a transgene carrying two copies of the normal human methylmalonyl-CoA mutase locus. The ‘‘humanized’’ mice were smaller than control litter mates and had high levels of methylmalonic acid in their blood and tissues. This new transgenic MMA stop codon model mimics (at both the phenotypic and genotypic levels) the key features of the human MMA disorder. It will allow the trialing of pharmacological and, cell and