Rosa26 Locus Supports Tissue-Specific Promoter Driving Transgene Expression Specifically in PigReportar como inadecuado




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Genetically modified pigs have become a popular model system in fundamental research, agricultural and biomedical applications. However, random integration often result in unstable expression of transgene and unpredictable phenotypes. The Rosa26 locus has been widely used to produce genetic modified animals with high and consistent expressing of transgene in mouse, human and rat, as it can be targeted efficiently and is not subject to gene-silencing effects. Recently, the first case of reporter gene targeting pigs in porcine Rosa26 pRosa26 locus was reported. In the study, full sequence of pRosa26 locus was further characterized, and the pRosa26 promoter pR26 was cloned and we evidenced that the new porcine endogenous promoter is suitable for driving transgene expression in a high and stable manner by avoiding DNA methylation. Furthermore, elongation factor 1a promoter EF1a -driven GFP reporter and Myostatin promoter MyoP-driven Follistatin Fst were successfully targeted into the pRosa26 locusby traditional homologous recombination HR strategy. EF1a showed high activity and hypomethylation at the locus. And, muscle-specific promoter MyoP was activated strictly in muscle of the pRosa26 targeted pigs, indicating Rosa26 locus supports tissue-specific promoter driving transgene expression in its own manner. The study provided further demonstration on biomedical and agricultural applications of porcine Rosa26 promoter and locus.



Autor: Qingran Kong , Tang Hai , Jing Ma, Tianqing Huang, Dandan Jiang, Bingteng Xie, Meiling Wu, Jiaqiang Wang, Yuran Song, Ying Wang,

Fuente: http://plos.srce.hr/



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