Establishment of a pig fibroblast-derived cell line for locus-directed transgene expression in cell cultures and blastocysts |
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| Authors: | Jannik E Jakobsen Juan Li Brian Moldt Peter M Kragh Henrik Callesen Jens Michael Hertz Lars Bolund Arne Lund Jørgensen Jacob Giehm Mikkelsen Anders Lade Nielsen |
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| Institution: | (1) Department of Human Genetics, Aarhus University, The Bartholin Building, 8000 Aarhus C, Denmark;(2) Section of Population Genetics and Embryology, Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, Aarhus University, 8830 Tjele, Denmark;(3) Department of Clinical Genetics, Aarhus University Hospital, Aarhus C, Denmark |
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| Abstract: | We report the establishment of a spontaneously immortalized pig cell line designated Pig Flip-in Visualize (PFV) for locus-directed
transgene expression in pig cells and blastocysts. The PFV cell line was isolated from pig ear fibroblasts transfected with
a Sleeping Beauty DNA transposon-based docking vector harbouring a selection gene, an eGFP reporter gene, and an Flp recombinase
site for locus-directed gene insertion. PFV cells have insertion of a single docking vector with stable eGFP expression and
generated phenotypic normal blastocysts with transgene expression after somatic cell nuclear transfer. PFV cells supported
Flp mediated cassette exchange for transgene substitution of eGFP with dsRED, and the dsRED transgenic PFV cells generated
blastocysts with transgene expression. Hence, the PFV cell line constitutes a valuable pig equivalent to transformed cell
lines from other mammalian species suitable for locus-directed transgene expression in cell cultures and, in addition, for
transgene analyses in the very early embryonic stages. |
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