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001-es BibID:BIBFORM059252
Első szerző:Garrels, Wiebke
Cím:Germline Transgenic Pigs by Sleeping Beauty Transposition in Porcine Zygotes and Targeted Integration in the Pig Genome / Wiebke Garrels, Lajos Mátés, Stephanie Holler, Anna Dalda, Ulrike Taylor, Björn Petersen, Heiner Niemann, Zsuzsanna Izsvák, Zoltán Ivics, Wilfried A. Kues
Dátum:2011
ISSN:1932-6203
Megjegyzések:Genetic engineering can expand the utility of pigs for modeling human diseases, and for developing advanced therapeutic approaches. However, the inefficient production of transgenic pigs represents a technological bottleneck. Here, we assessed the hyperactive Sleeping Beauty (SB100X) transposon system for enzyme-catalyzed transgene integration into the embryonic porcine genome. The components of the transposon vector system were microinjected as circular plasmids into the cytoplasm of porcine zygotes, resulting in high frequencies of transgenic fetuses and piglets. The transgenic animals showed normal development and persistent reporter gene expression for >12 months. Molecular hallmarks of transposition were confirmed by analysis of 25 genomic insertion sites. We demonstrate germ-line transmission, segregation of individual transposons, and continued, copy number-dependent transgene expression in F1-offspring. In addition, we demonstrate target-selected gene insertion into transposon-tagged genomic loci by Cre-loxP-based cassette exchange in somatic cells followed by nuclear transfer. Transposase-catalyzed transgenesis in a large mammalian species expands the arsenal of transgenic technologies for use in domestic animals and will facilitate the development of large animal models for human diseases.
Tárgyszavak:Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
transposable elements
swine
animal genomics
fetuses
fibroblasts
zygotes
reporter gene assay
genetically modified animals
Megjelenés:Plos One. - 6 : 8 (2011), p. e23573-. -
További szerzők:Mátés Lajos Holler, Stephanie Dalda Anna Taylor, Ulrike Petersen, Björn Niemann, Heiner Izsvák Zsuzsanna (1961-) (genetikus) Ivics Zoltán (1964-) (biológus) Kues, Wilfried A.
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