Generation and characterization of transgenic mice with the full-length human DMD gene.

摘要: We report the generation of mice with an intact and functional copy 2.3-megabase human dystrophin gene (hDMD), largest stretch DNA thus far integrated into a mouse chromosome. Yeast spheroplasts containing artificial chromosome full-length hDMD were fused embryonic stem cells subsequently injected blastocysts to produce transgenic mice. Human-specific PCR, Southern blotting, fluorescent in situ hybridization techniques demonstrated intactness stable chromosomal integration on 5. Expression transgene was confirmed by RT-PCR Western blotting. The tissue-specific expression pattern different DMD transcripts maintained. However, Dp427p Dp427m expressed at 2-fold higher levels Dp427c Dp260 2- 4-fold lower than their endogenous counterparts. Ultimate proof obtained crossing dystrophin-deficient mdx utrophin-deficient × Utrn-/- rescued lethal dystrophic phenotype All signs muscular dystrophy disappeared mice, as histological staining muscle sections profiling experiments. Currently, are extensively used for preclinical testing sequence-specific therapeutics treatment Duchenne dystrophy. In addition, can be study influence genomic context deletion recombination frequencies, genome stability, regulation.