Generation of interleukin-2 receptor gamma gene knockout pigs from somatic cells genetically modified by zinc finger nuclease-encoding mRNA.
Zinc finger nuclease (ZFN) is a powerful tool for genome editing. ZFN-encoding plasmid DNA expression systems have been recently employed for the generation of gene knockout (KO) pigs, although one major limitation of this technology is the use of potentially harmful genome-integrating plasmid DNAs....
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3793986?pdf=render |
| _version_ | 1818826410276421632 |
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| author | Masahito Watanabe Kazuaki Nakano Hitomi Matsunari Taisuke Matsuda Miki Maehara Takahiro Kanai Mirina Kobayashi Yukina Matsumura Rieko Sakai Momoko Kuramoto Gota Hayashida Yoshinori Asano Shuko Takayanagi Yoshikazu Arai Kazuhiro Umeyama Masaki Nagaya Yutaka Hanazono Hiroshi Nagashima |
| author_facet | Masahito Watanabe Kazuaki Nakano Hitomi Matsunari Taisuke Matsuda Miki Maehara Takahiro Kanai Mirina Kobayashi Yukina Matsumura Rieko Sakai Momoko Kuramoto Gota Hayashida Yoshinori Asano Shuko Takayanagi Yoshikazu Arai Kazuhiro Umeyama Masaki Nagaya Yutaka Hanazono Hiroshi Nagashima |
| author_sort | Masahito Watanabe |
| collection | DOAJ |
| description | Zinc finger nuclease (ZFN) is a powerful tool for genome editing. ZFN-encoding plasmid DNA expression systems have been recently employed for the generation of gene knockout (KO) pigs, although one major limitation of this technology is the use of potentially harmful genome-integrating plasmid DNAs. Here we describe a simple, non-integrating strategy for generating KO pigs using ZFN-encoding mRNA. The interleukin-2 receptor gamma (IL2RG) gene was knocked out in porcine fetal fibroblasts using ZFN-encoding mRNAs, and IL2RG KO pigs were subsequently generated using these KO cells through somatic cell nuclear transfer (SCNT). The resulting IL2RG KO pigs completely lacked a thymus and were deficient in T and NK cells, similar to human X-linked SCID patients. Our findings demonstrate that the combination of ZFN-encoding mRNAs and SCNT provides a simple robust method for producing KO pigs without genomic integration. |
| first_indexed | 2024-12-19T00:27:13Z |
| format | Article |
| id | doaj.art-8b3107db5c99480d9579f29e3686a755 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-19T00:27:13Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-8b3107db5c99480d9579f29e3686a7552022-12-21T20:45:14ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-01810e7647810.1371/journal.pone.0076478Generation of interleukin-2 receptor gamma gene knockout pigs from somatic cells genetically modified by zinc finger nuclease-encoding mRNA.Masahito WatanabeKazuaki NakanoHitomi MatsunariTaisuke MatsudaMiki MaeharaTakahiro KanaiMirina KobayashiYukina MatsumuraRieko SakaiMomoko KuramotoGota HayashidaYoshinori AsanoShuko TakayanagiYoshikazu AraiKazuhiro UmeyamaMasaki NagayaYutaka HanazonoHiroshi NagashimaZinc finger nuclease (ZFN) is a powerful tool for genome editing. ZFN-encoding plasmid DNA expression systems have been recently employed for the generation of gene knockout (KO) pigs, although one major limitation of this technology is the use of potentially harmful genome-integrating plasmid DNAs. Here we describe a simple, non-integrating strategy for generating KO pigs using ZFN-encoding mRNA. The interleukin-2 receptor gamma (IL2RG) gene was knocked out in porcine fetal fibroblasts using ZFN-encoding mRNAs, and IL2RG KO pigs were subsequently generated using these KO cells through somatic cell nuclear transfer (SCNT). The resulting IL2RG KO pigs completely lacked a thymus and were deficient in T and NK cells, similar to human X-linked SCID patients. Our findings demonstrate that the combination of ZFN-encoding mRNAs and SCNT provides a simple robust method for producing KO pigs without genomic integration.http://europepmc.org/articles/PMC3793986?pdf=render |
| spellingShingle | Masahito Watanabe Kazuaki Nakano Hitomi Matsunari Taisuke Matsuda Miki Maehara Takahiro Kanai Mirina Kobayashi Yukina Matsumura Rieko Sakai Momoko Kuramoto Gota Hayashida Yoshinori Asano Shuko Takayanagi Yoshikazu Arai Kazuhiro Umeyama Masaki Nagaya Yutaka Hanazono Hiroshi Nagashima Generation of interleukin-2 receptor gamma gene knockout pigs from somatic cells genetically modified by zinc finger nuclease-encoding mRNA. PLoS ONE |
| title | Generation of interleukin-2 receptor gamma gene knockout pigs from somatic cells genetically modified by zinc finger nuclease-encoding mRNA. |
| title_full | Generation of interleukin-2 receptor gamma gene knockout pigs from somatic cells genetically modified by zinc finger nuclease-encoding mRNA. |
| title_fullStr | Generation of interleukin-2 receptor gamma gene knockout pigs from somatic cells genetically modified by zinc finger nuclease-encoding mRNA. |
| title_full_unstemmed | Generation of interleukin-2 receptor gamma gene knockout pigs from somatic cells genetically modified by zinc finger nuclease-encoding mRNA. |
| title_short | Generation of interleukin-2 receptor gamma gene knockout pigs from somatic cells genetically modified by zinc finger nuclease-encoding mRNA. |
| title_sort | generation of interleukin 2 receptor gamma gene knockout pigs from somatic cells genetically modified by zinc finger nuclease encoding mrna |
| url | http://europepmc.org/articles/PMC3793986?pdf=render |
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