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

Disease Models & Mechanisms
Ana Isabel Garcia DiazTim Aitman

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 exte...Continue Reading

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Citations

Oct 5, 2016·Critical Reviews in Biochemistry and Molecular Biology·Suneel A NarayanavariZsuzsanna Izsvák
Dec 10, 2016·Disease Models & Mechanisms·Linda J MullinsJohn J Mullins
Jul 26, 2019·Physiological Research·N LiptákL Hiripi
Sep 2, 2020·Journal of the American Society of Nephrology : JASN·Tabitha Turner-StokesKevin J Woollard
Jul 20, 2019·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Rumyana SimeonovaReneta Gevrenova
Jan 29, 2021·International Journal of Molecular Sciences·Natalie VallantVassilios Papalois

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Methods Mentioned

BETA
transgenic
PCR
fluorescence microscopy
flow cytometry
dissection
dissections
confocal microscopy

Software Mentioned

ARRIVE
Rat Rnor
Ensembl browser

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