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New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein

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-estab...

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Autores principales: Garcia Diaz, Ana Isabel, Moyon, Ben, Coan, Philip M., Alfazema, Neza, Venda, Lara, Woollard, Kevin, Aitman, Tim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Company of Biologists Ltd 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4852507/
https://www.ncbi.nlm.nih.gov/pubmed/26769799
http://dx.doi.org/10.1242/dmm.024208
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author Garcia Diaz, Ana Isabel
Moyon, Ben
Coan, Philip M.
Alfazema, Neza
Venda, Lara
Woollard, Kevin
Aitman, Tim
author_facet Garcia Diaz, Ana Isabel
Moyon, Ben
Coan, Philip M.
Alfazema, Neza
Venda, Lara
Woollard, Kevin
Aitman, Tim
author_sort Garcia Diaz, Ana Isabel
collection PubMed
description 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 preliminary in vitro and in vivo imaging 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.
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spelling pubmed-48525072016-05-19 New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein Garcia Diaz, Ana Isabel Moyon, Ben Coan, Philip M. Alfazema, Neza Venda, Lara Woollard, Kevin Aitman, Tim Dis Model Mech Resource Article 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 preliminary in vitro and in vivo imaging 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. The Company of Biologists Ltd 2016-04-01 /pmc/articles/PMC4852507/ /pubmed/26769799 http://dx.doi.org/10.1242/dmm.024208 Text en © 2016. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
spellingShingle Resource Article
Garcia Diaz, Ana Isabel
Moyon, Ben
Coan, Philip M.
Alfazema, Neza
Venda, Lara
Woollard, Kevin
Aitman, Tim
New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein
title New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein
title_full New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein
title_fullStr New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein
title_full_unstemmed New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein
title_short New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein
title_sort new wistar kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein
topic Resource Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4852507/
https://www.ncbi.nlm.nih.gov/pubmed/26769799
http://dx.doi.org/10.1242/dmm.024208
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