title: Rosa26 locus supports tissue-specific promoter driving transgene expression specifically in pig.
abstract: 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.
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Qingran
authors.initials: Q
authors.lastname: Kong
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Tang
authors.initials: T
authors.lastname: Hai
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Ma
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Tianqing
authors.initials: T
authors.lastname: Huang
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Jiang
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Bingteng
authors.initials: B
authors.lastname: Xie
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Meiling
authors.initials: M
authors.lastname: Wu
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Jiaqiang
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China; Graduate University of the Chinese Academy of Sciences, Beijing, China.
authors.forename: Yuran
authors.initials: Y
authors.lastname: Song
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Yilong
authors.initials: Y
authors.lastname: He
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Jialu
authors.initials: J
authors.lastname: Sun
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Kui
authors.initials: K
authors.lastname: Hu
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Runfa
authors.initials: R
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Liu
authors.initials: L
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Yanshuang
authors.initials: Y
authors.lastname: Mu
authors.affiliation: Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin, China.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Liu
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D002478:Cells, Cultured; D015870:Gene Expression; D017930:Genes, Reporter; D056426:Genetic Loci; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D022661:RNA, Untranslated / Q000235:genetics*; D034421:Sus scrofa / Q000235:genetics*; D013552:Swine; D013556:Swine, Miniature; D015533:Transcriptional Activation
chemical_list: D022661:RNA, Untranslated; D049452:Green Fluorescent Proteins