2003 Fiscal Year Final Research Report Summary
Establishment of prion deficient cells in cattle
| Project/Area Number |
14560281
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物資源科学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
SHINKAI Yoichi Kyoto University, Institute for Virus Research, Professor, ウイルス研究所, 教授 (20211972)
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| Co-Investigator(Kenkyū-buntansha) |
SENDAI Yutaka Kyoto University, Research Institute for Functional Peptides, Senior Researcher, 研究員
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| Project Period (FY) |
2002 – 2003
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| Keywords | PRION / CATTLE / GENE TARGETING / BSE |
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| Research Abstract |
Animal cloning techniques have enabled gene disruption in several species. Here, we report the first successful disruption of the prion gene in cattle. METHODS : The prion gene of the Japanese Black cow (JBC) was used to construct PrP-5. 1 and PrP-5. 2, the targeting vectors for the bovine prion gene, and these constructs were introduced into the fetal fibroblast cell line JBC906 by the lipofection method. Ten polymerase chain reaction (PCR)-positive colonies were obtained from 833 G418-resistant colonies, and Southern blot analysis revealed successful homologous recombination at the prion locus in these colonies. CONCLUSIONS : Heterozygous knockout of the prion gene was performed in JBC, and production of a homozygous prion knockout JBC by a second round of targeting 906htGT is currently in progress.
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