Establishment of a pig fibroblast-derived cell line for locus-directed transgene expression in cell cultures and blastocysts
Establishment of a pig fibroblast-derived cell line for locus-directed transgene expression in cell cultures and blastocysts
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DOI：
10.1007/s11033-010-0089-z
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发表时间：
2011-01-01
影响因子：
2.8
通讯作者：
Nielsen, Anders Lade
中科院分区：
文献类型：
--
作者：
Jakobsen, Jannik E.;Li, Juan;Nielsen, Anders Lade
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.
