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Identification and re-addressing of a transcriptionally permissive locus in the porcine genome

Item Type:Article
Title:Identification and re-addressing of a transcriptionally permissive locus in the porcine genome
Creators Name:Garrels, W. and Mukherjee, A. and Holler, S. and Cleve, N. and Talluri, T.R. and Barg-Kues, B. and Diederich, M. and Köhler, P. and Petersen, B. and Lucas-Hahn, A. and Niemann, H. and Izsvák, Z. and Ivics, Z. and Kues, W.A.
Abstract:Recently, we established the Sleeping Beauty transposon system for germ line competent transgenesis in the pig. Here, we extend this approach to re-target a transposon-tagged locus for a site-specific gene knock-in, and generated a syngeneic cohort of piglets carrying either the original transposon or the re-targeted event. A Cre-loxP-mediated cassette exchange of the tagging transposon with a different reporter gene was performed, followed by flow cytometric sorting and somatic cell nuclear transfer of recombined cells. In parallel, the original cells were employed in somatic cell nuclear transfer to generate clone siblings, thereby resulting in a clone cohort of piglets carrying different reporter transposons at an identical chromosomal location. Importantly, this strategy supersedes the need for an antibiotic selection marker. This approach expands the arsenal of genome engineering technologies in domestic animals, and will facilitate the development of large animal models for human diseases. Potentially, the syngeneic cohort of pigs will be instrumental for vital tracking of transplanted cells in pre-clinical assessments of novel cell therapies.
Keywords:Large Animal Model, Active Transgenesis, Targeted Integration, Somatic Cell Nuclear Transfer, Livestock, Animals, Sus scrofa
Source:Transgenic Research
ISSN:0962-8819
Publisher:Springer (Netherlands)
Volume:25
Number:1
Page Range:63-70
Date:February 2016
Official Publication:https://doi.org/10.1007/s11248-015-9914-4
PubMed:View item in PubMed

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