New Wistar Kyoto and Spontaneously Hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein

Autor: Timothy J. Aitman, Ana-Isabel Garcia Diaz, Kevin J. Woollard, Lara Venda, Ben Moyon, Neza Alfazema, P. M. Coan
Přispěvatelé: Nan Dimond Charitable Trust
Rok vydání: 2016
Předmět:
0301 basic medicine
Knockout rat
Intravital Microscopy
Gene Expression
lcsh:Medicine
Medicine (miscellaneous)
SLEEPING-BEAUTY TRANSPOSONS
Rats
Inbred WKY
Green fluorescent protein
SHR
Immunology and Microbiology (miscellaneous)
ANIMAL-MODEL
Rats
Inbred SHR
Gene expression
Pathology
Leukocytes
GENE-EXPRESSION
Genetics
Gene Transfer Techniques
11 Medical And Health Sciences
Transgenesis
Organ Specificity
Models
Animal
Rats
Transgenic
CRESCENTIC GLOMERULONEPHRITIS
Life Sciences & Biomedicine
lcsh:RB1-214
Microinjections
Transgene
Green Fluorescent Proteins
Neuroscience (miscellaneous)
Bone Marrow Cells
Biology
GFP
EMBRYO MICROINJECTION
General Biochemistry
Genetics and Molecular Biology
03 medical and health sciences
Spontaneously hypertensive rat
lcsh:Pathology
Animals
Resource Article
LENTIVIRAL VECTORS
Science & Technology
KNOCKOUT RATS
FATTY-ACID
IDENTIFICATION
HUMAN-CELLS
Macrophages
lcsh:R
Cell Biology
06 Biological Sciences
Embryo
Mammalian
Sleeping Beauty transposon system
Molecular biology
Transplantation
030104 developmental biology
DNA Transposable Elements
WKY
Rat
Developmental Biology
Transgenics
Zdroj: Disease Models & Mechanisms, Vol 9, Iss 4, Pp 463-471 (2016)
Disease Models & Mechanisms
Diaz, A-I G, Moyon, B, Coan, P, Alfazema, N, Venda, L, Woollard, K & Aitman, T 2016, ' New Wistar Kyoto and Spontaneously Hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein ', Disease Models and Mechanisms . https://doi.org/10.1242/dmm.024208
ISSN: 1754-8411
1754-8403
Popis: 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.
Summary: We have created two transgenic models expressing ubiquitous GFP in WKY and SHR rat lines to support in vivo and ex vivo studies. We show their characterisation and discuss their potential use for translational research.
Databáze: OpenAIRE
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