Generation and characterization of a transgenic pig carrying a DsRed-Monomer reporter gene
Journal
PLoS ONE
Journal Volume
9
Journal Issue
9
Date Issued
2014
Author(s)
Abstract
Background: Pigs are an optimal animal for conducting biomedical research because of their anatomical and physiological resemblance to humans. In contrast to the abundant resources available in the study of mice, few fluorescent protein-harboring porcine models are available for preclinical studies. In this paper, we report the successful generation and characterization of a transgenic DsRed-Monomer porcine model. Methods: The transgene comprised a CMV enhancer/chicken-beta actin promoter and DsRed monomeric cDNA. Transgenic pigs were produced by using pronuclear microinjection. PCR and Southern blot analyses were applied for identification of the transgene. Histology, blood examinations and computed tomography were performed to study the health conditions. The pig amniotic fluid progenitor/stem cells were also isolated to examine the existence of red fluorescence and differentiation ability. Results: Transgenic pigs were successfully generated and transmitted to offspring at a germ-line transmission rate of 43.59% (17/39). Ubiquitous expression of red fluorescence was detected in the brain, eye, tongue, heart, lung, liver, pancreas, spleen, stomach, small intestine, large intestine, kidney, testis, and muscle; this was confirmed by histology and western blot analyses. In addition, we confirmed the differentiation potential of amniotic fluid progenitor stem cells isolated from the transgenic pig. Conclusions: This red fluorescent pig can serve as a host for other fluorescent-labeled cells in order to study cell-microenvironment interactions, and can provide optimal red-fluorescent-labeled cells and tissues for research in developmental biology, regenerative medicine, and xenotransplantation. ? 2014 Chou et al.
SDGs
Other Subjects
red fluorescent protein; actin; complementary DNA; fluorescent protein 583; photoprotein; amniotic fluid cell; animal cell; animal experiment; animal tissue; Article; blood examination; brain; cell differentiation; cell isolation; cellular distribution; computer assisted tomography; controlled study; DsRed gene; eye; genetic identification; germline mutation; heart; histology; kidney; liver; lung; muscle; nonhuman; pancreas; polymerase chain reaction; protein expression; reporter gene; small intestine; Southern blotting; spleen; stomach; testis; tongue; transgenic pig; Western blotting; amnion fluid; animal; cell culture; chemistry; chicken; cytology; Cytomegalovirus; embryo transfer; enhancer region; female; fluorescence; founder effect; gene expression; genetics; male; metabolism; microinjection; physiology; pig; promoter region; reporter gene; stem cell; transgenic animal; veterinary; Suidae; Actins; Amniotic Fluid; Animals; Animals, Genetically Modified; Cell Differentiation; Cells, Cultured; Chickens; Cytomegalovirus; DNA, Complementary; Enhancer Elements, Genetic; Female; Fluorescence; Founder Effect; Gene Expression; Genes, Reporter; Luminescent Proteins; Male; Microinjections; Promoter Regions, Genetic; Stem Cells; Swine; Zygote Intrafallopian Transfer
Type
journal article