Establishment of a transgenic pig fetal fibroblast reporter cell line for monitoring Cre recombinase activity

• Published in: DNA and cell biology Vol. 28; no. 6; p. 303
• Main Authors: Li, Li, Pang, Daxin, Chen, Limei, Wang, Tiedong, Nie, Daibang, Yan, Sen, Ouyang, Hongsheng
• Format: Journal Article
• Language: English
• Published: United States 01.06.2009
• Subjects: Animals; Animals, Genetically Modified; Cell Line; Fetus - cytology; Fibroblasts - metabolism; Flow Cytometry; Gene Knockout Techniques; Genes, Reporter; Genes, Synthetic; Green Fluorescent Proteins - analysis; Green Fluorescent Proteins - genetics; Humans; Integrases - analysis; Microscopy, Confocal; Recombinant Fusion Proteins - analysis; Recombinant Fusion Proteins - genetics; Reverse Transcriptase Polymerase Chain Reaction; Sus scrofa - embryology; Sus scrofa - genetics; Transfection; Transgenes
• ISSN: 1557-7430
• DOI: 10.1089/dna.2008.0821

The pig is considered to be the most suitable nonhuman source of organs for xenotransplantation and is widely used as a model of human disease. The Cre-LoxP system provides a powerful means of cell- or tissue-specific deletion of a targeted gene in cells or tissues of interest. Pigs expressing Cre recombinase have a profound impact on the study of gene function and the generation of animal models of human diseases. To monitor Cre recombinase expression in vivo, it is important to create reporter strains. As a first step in the production of such transgenic pigs, we generated porcine fetal fibroblast cell lines conditionally expressing the gene for enhanced green fluorescent protein (EGFP). The EGFP gene is expressed only after Cre-mediated excision of LoxP-flanked stop sequences. These fetal fibroblast cell lines will be of great value for constructing reporter transgenic pigs.