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PLoS One. 2013;8(2):e56518. doi: 10.1371/journal.pone.0056518. Epub 2013 Feb 20.

Derivation of a germline competent transgenic Fischer 344 embryonic stem cell line.

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1
Rat Resource and Research Center, Department of Veterinary Pathobiology, University of Missouri, Columbia, Missouri, USA.

Abstract

Embryonic stem (ES) cell-based gene manipulation is an effective method for the generation of mutant animal models in mice and rats. Availability of germline-competent ES cell lines from inbred rat strains would allow for creation of new genetically modified models in the desired genetic background. Fischer344 (F344) males carrying an enhanced green fluorescence protein (EGFP) transgene were used as the founder animals for the derivation of ES cell lines. After establishment of ES cell lines, rigorous quality control testing that included assessment of pluripotency factor expression, karyotype analysis, and pathogen/sterility testing was conducted in selected ES cell lines. One male ES cell line, F344-Tg.EC4011, was further evaluated for germline competence by injection into Dark Agouti (DA) X Sprague Dawley (SD) blastocysts. Resulting chimeric animals were bred with wild-type SD mates and germline transmissibility of the ES cell line was confirmed by identification of pups carrying the ES cell line-derived EGFP transgene. This is the first report of a germline competent F344 ES cell line. The availability of a new germline competent ES cell line with a stable fluorescence reporter from an inbred transgenic rat strain provides an important new resource for genetic manipulations to create new rat models.

PMID:
23437152
PMCID:
PMC3577902
DOI:
10.1371/journal.pone.0056518
[Indexed for MEDLINE]
Free PMC Article
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