title: New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein.
abstract: The Wistar Kyoto (WKY) rat and the spontaneously hypertensive (SHR) rat inbred strains are well-established models for human crescentic glomerulonephritis (CRGN) and metabolic syndrome, respectively. Novel transgenic (Tg) strains add research opportunities and increase scientific value to well-established rat models. We have created two novel Tg strains using Sleeping Beauty transposon germline transgenesis, ubiquitously expressing green fluorescent protein (GFP) under the rat elongation factor 1 alpha (EF1a) promoter on the WKY and SHR genetic backgrounds. The Sleeping Beauty system functioned with high transgenesis efficiency; 75% of new rats born after embryo microinjections were transgene positive. By ligation-mediated PCR, we located the genome integration sites, confirming no exonic disruption and defining a single or low copy number of the transgenes in the new WKY-GFP and SHR-GFP Tg lines. We report GFP-bright expression in embryos, tissues and organs in both lines and show preliminaryin vitroandin vivoimaging data that demonstrate the utility of the new GFP-expressing lines for adoptive transfer, transplantation and fate mapping studies of CRGN, metabolic syndrome and other traits for which these strains have been extensively studied over the past four decades.
authors.affiliation: Division of Immunology and Inflammation, Imperial College London, London W2 1PG, UK MRC Clinical Sciences Centre and Department of Medicine, Imperial College London, London W12 0NN, UK.
authors.forename: Ana Isabel
authors.initials: AI
authors.lastname: Garcia Diaz
authors.affiliation: Embryonic Stem Cell and Transgenics Facility, MRC Clinical Sciences Centre, Imperial College London, London W12 0NN, UK.
authors.forename: Ben
authors.initials: B
authors.lastname: Moyon
authors.affiliation: Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh EH4 2XU, UK.
authors.forename: Philip M
authors.initials: PM
authors.lastname: Coan
authors.affiliation: Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh EH4 2XU, UK.
authors.forename: Neza
authors.initials: N
authors.lastname: Alfazema
authors.affiliation: MRC Clinical Sciences Centre and Department of Medicine, Imperial College London, London W12 0NN, UK.
authors.forename: Lara
authors.initials: L
authors.lastname: Venda
authors.affiliation: Division of Immunology and Inflammation, Imperial College London, London W2 1PG, UK k.woollard@imperial.ac.uk tim.aitman@ed.ac.uk.
authors.forename: Kevin
authors.initials: K
authors.lastname: Woollard
authors.affiliation: MRC Clinical Sciences Centre and Department of Medicine, Imperial College London, London W12 0NN, UK Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh EH4 2XU, UK k.woollard@imperial.ac.uk tim.aitman@ed.ac.uk.
authors.forename: Tim
authors.initials: T
authors.lastname: Aitman
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D004251:DNA Transposable Elements / Q000235:genetics; D004622:Embryo, Mammalian / Q000378:metabolism; D015870:Gene Expression; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000097:blood / Q000235:genetics*; D000069416:Intravital Microscopy; D007962:Leukocytes / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D008845:Microinjections; D023421:Models, Animal; D009928:Organ Specificity; D011918:Rats, Inbred SHR; D011921:Rats, Inbred WKY; D055647:Rats, Transgenic
chemical_list: D004251:DNA Transposable Elements; D049452:Green Fluorescent Proteins
keywords: GFP; Rat; SHR; Transgenics; WKY