Establishment of prion deficient cells in cattle

牛朊病毒缺陷细胞的建立

• 批准号: 14560281
• 负责人: SHINKAI Yoichi
• 金额: $ 2.75万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14560281/
• 关键词: PRION; CATTLE; GENE TARGETING; BSE; クローニング; ノックアウト

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.

动物克隆技术已经使一些物种的基因受到破坏。在这里，我们报告了首次成功破坏牛的朊病毒基因。方法：采用日本黑牛（JBC）朊病毒基因构建PrP-5。1和PrP-5。2、将牛朊病毒基因的靶向载体和这些构建物通过脂质转染法导入胎成纤维细胞系JBC906。从833个g418抗性菌落中获得10个聚合酶链反应（PCR）阳性菌落，经Southern blot分析，这些菌落的朊病毒位点同源重组成功。结论：在JBC中进行了朊病毒基因的杂合子敲除，目前正在进行第二轮靶向906htGT的纯合子朊病毒敲除JBC。