title: Transposon-mediated transgenesis, transgenic rescue, and tissue-specific gene expression in rodents and rabbits.
abstract: Germline transgenesis is an important procedure for functional investigation of biological pathways, as well as for animal biotechnology. We have established a simple, nonviral protocol in three important biomedical model organisms frequently used in physiological studies. The protocol is based on the hyperactive Sleeping Beauty transposon system, SB100X, which reproducibly promoted generation of transgenic founders at frequencies of 50-64, 14-72, and 15% in mice, rats, and rabbits, respectively. The SB100X-mediated transgene integrations are less prone to genetic mosaicism and gene silencing as compared to either the classical pronuclear injection or to lentivirus-mediated transgenesis. The method was successfully applied to a variety of transgenes and animal models, and can be used to generate founders with single-copy integrations. The transposon vector also allows the generation of transgenic lines with tissue-specific expression patterns specified by promoter elements of choice, exemplified by a rat reporter strain useful for tracking serotonergic neurons. As a proof of principle, we rescued an inborn genetic defect in the fawn-hooded hypertensive rat by SB100X transgenesis. A side-by-side comparison of the SB100X- and piggyBac-based protocols revealed that the two systems are complementary, offering new opportunities in genome manipulation.
authors.affiliation: Institute of Laboratory Animal Science, University of Veterinary Medicine Vienna, Vienna, Austria.
authors.forename: Katharina
authors.initials: K
authors.lastname: Katter
authors.forename: Aron M
authors.initials: AM
authors.lastname: Geurts
authors.forename: Orsolya
authors.initials: O
authors.lastname: Hoffmann
authors.forename: Lajos
authors.initials: L
authors.lastname: Mátés
authors.forename: Vladimir
authors.initials: V
authors.lastname: Landa
authors.forename: László
authors.initials: L
authors.lastname: Hiripi
authors.forename: Carol
authors.initials: C
authors.lastname: Moreno
authors.forename: Jozef
authors.initials: J
authors.lastname: Lazar
authors.forename: Sanum
authors.initials: S
authors.lastname: Bashir
authors.forename: Vaclav
authors.initials: V
authors.lastname: Zidek
authors.forename: Elena
authors.initials: E
authors.lastname: Popova
authors.forename: Boris
authors.initials: B
authors.lastname: Jerchow
authors.forename: Katja
authors.initials: K
authors.lastname: Becker
authors.forename: Anantharam
authors.initials: A
authors.lastname: Devaraj
authors.forename: Ingrid
authors.initials: I
authors.lastname: Walter
authors.forename: Michael
authors.initials: M
authors.lastname: Grzybowksi
authors.forename: Molly
authors.initials: M
authors.lastname: Corbett
authors.forename: Artur Rangel
authors.initials: AR
authors.lastname: Filho
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Hodges
authors.forename: Michael
authors.initials: M
authors.lastname: Bader
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Ivics
authors.forename: Howard J
authors.initials: HJ
authors.lastname: Jacob
authors.forename: Michal
authors.initials: M
authors.lastname: Pravenec
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Bosze
authors.forename: Thomas
authors.initials: T
authors.lastname: Rülicke
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Izsvák
mesh_terms: D000818:Animals; D004251:DNA Transposable Elements / Q000235:genetics*; D005260:Female; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009030:Mosaicism; D009928:Organ Specificity / Q000235:genetics; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D019076:Transgenes
chemical_list: D004251:DNA Transposable Elements