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Generation and Characterization of a Transgenic Pig Carrying a DsRed-Monomer Reporter Gene

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....

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Autores principales: Chou, Chih-Jen, Peng, Shao-Yu, Wu, Mei-Han, Yang, Cho-Chen, Lin, Yu-Sheng, Cheng, Winston Teng-Kui, Wu, Shinn-Chih, Lin, Yao-Ping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4154781/
https://www.ncbi.nlm.nih.gov/pubmed/25187950
http://dx.doi.org/10.1371/journal.pone.0106864
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author Chou, Chih-Jen
Peng, Shao-Yu
Wu, Mei-Han
Yang, Cho-Chen
Lin, Yu-Sheng
Cheng, Winston Teng-Kui
Wu, Shinn-Chih
Lin, Yao-Ping
author_facet Chou, Chih-Jen
Peng, Shao-Yu
Wu, Mei-Han
Yang, Cho-Chen
Lin, Yu-Sheng
Cheng, Winston Teng-Kui
Wu, Shinn-Chih
Lin, Yao-Ping
author_sort Chou, Chih-Jen
collection PubMed
description 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.
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spelling pubmed-41547812014-09-08 Generation and Characterization of a Transgenic Pig Carrying a DsRed-Monomer Reporter Gene Chou, Chih-Jen Peng, Shao-Yu Wu, Mei-Han Yang, Cho-Chen Lin, Yu-Sheng Cheng, Winston Teng-Kui Wu, Shinn-Chih Lin, Yao-Ping PLoS One Research Article 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. Public Library of Science 2014-09-04 /pmc/articles/PMC4154781/ /pubmed/25187950 http://dx.doi.org/10.1371/journal.pone.0106864 Text en © 2014 Chou et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Chou, Chih-Jen
Peng, Shao-Yu
Wu, Mei-Han
Yang, Cho-Chen
Lin, Yu-Sheng
Cheng, Winston Teng-Kui
Wu, Shinn-Chih
Lin, Yao-Ping
Generation and Characterization of a Transgenic Pig Carrying a DsRed-Monomer Reporter Gene
title Generation and Characterization of a Transgenic Pig Carrying a DsRed-Monomer Reporter Gene
title_full Generation and Characterization of a Transgenic Pig Carrying a DsRed-Monomer Reporter Gene
title_fullStr Generation and Characterization of a Transgenic Pig Carrying a DsRed-Monomer Reporter Gene
title_full_unstemmed Generation and Characterization of a Transgenic Pig Carrying a DsRed-Monomer Reporter Gene
title_short Generation and Characterization of a Transgenic Pig Carrying a DsRed-Monomer Reporter Gene
title_sort generation and characterization of a transgenic pig carrying a dsred-monomer reporter gene
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4154781/
https://www.ncbi.nlm.nih.gov/pubmed/25187950
http://dx.doi.org/10.1371/journal.pone.0106864
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