Production of cloned genetically modified pigs by a new drug-free gene-modified cell isolation system
| Project/Area Number |
24580411
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Applied animal science
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| Research Institution | Kagoshima University |
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Principal Investigator |
SATO Masahiro 鹿児島大学, 医用ミニブタ・先端医療開発研究センター, 教授 (30287099)
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| Co-Investigator(Kenkyū-buntansha) |
MIYOSHI Kazuchika 鹿児島大学農学部, 生物生産学科, 教授 (70363611)
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Keywords | クローンブタ / 遺伝子導入 / ＫＯ / 遺伝子改変 / 異種移植 / DAF（補体抑制因子） / thrombomodulin（血液凝固阻害因子） / α-Gal epitope / 異種移植拒絶関連遺伝子 / ブタ細胞 / α-Galエピトープ / 体細胞核移植 / EndoGalC / CD47 / DAF / トロンボモジュリン |
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| Outline of Final Research Achievements |
Our purpose of this study is to produce genetically modified piglets that are used for xenotransplantation. α-GalT is known to synthesize α-Gal epitope (α-Gal), which is a causative for hyperacute rejection of pig grafts. We first disrupted the α-GalT gene in porcine cells using a recently developed technology that allows efficient ablation of a target gene, and finally obtained a cell line called CRISPR-2-1 (2-1). DAF and TM are also known as key factors suppressing graft rejection. In order to deliver genes for these two factors into 2-1, we developed a novel system that does not require drug selection and also employed a piggyBac-based system that permits introduction of multiple transgenes. We are now constructing 2-1-based new cell lines that are able to express both DAF and TM in the absence of α-Gal. The resulting cell line will be used for SCNT as donors to produce cloned piglets capable of expressing DAF and TM, but unable to express α-Gal.
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Report
(4 results)
Research Products
(52 results)
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[Journal Article] Identification of genomic locus responsible for experimentally induced testicular teratoma 1 (ett1) on mouse Chr 182014
Author(s)
Miyazaki T, Ikeda Y, Kubo I, Suganuma S, Fujita N, Itakura M, Hayashi T, Takabayashi S, Katoh H, Ohira Y, Sato M, Noguchi M, Tokumoto T
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Journal Title
Mammalian Genome
Volume: 25
Issue: 7-8
Pages: 317-326
DOI
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Peer Reviewed / Open Access
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[Journal Article] CD81 and CD9 work independently as extracellular components upon fusion of sperm and oocyt2012
Author(s)
Ohnami N, Nakamura A, Miyado M, Sato M, Kawano N, Yoshida K, Harada Y, Takezawa Y, Kanai S, Ono C, Takahashi Y, Kimura K, Shida T, Miyado K, Umezawa A
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Journal Title
Biol Open
Volume: 1(7)
Issue: 7
Pages: 640-647
DOI
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Peer Reviewed
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[Journal Article] Targeted transgenesis through pronuclear injection of improved vectors into in vitro fertilized eggs2011
Author(s)
Ohtsuka, M., Miura, H., Nakaoka, H., Kimura, M., Sato, M., Inoko, H.
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Journal Title
Transgenic Research
Volume: 21:1
Issue: 1
Pages: 225-226
DOI
Related Report
Peer Reviewed
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[Presentation] Efficient generation of α-1,3-galactosyltransferase-deficient porcine embryonic fibroblasts by CRISPR/Cas9-mediated knock-in system2014
Author(s)
Sato M, Kagoshima A, Saitoh I, Inada E, Miyoshi K, Ohtsuka M, Nakamura S, Sakurai T, Watanabe S
Organizer
The Asian Federation of Laboratory Animal Science Associations (AFLAS) Congress
Place of Presentation
Loyal Chulan Hotel, Kuala Lumpur, Malaysia
Year and Date
2014-11-11 – 2014-11-12
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