Efficient PRNP gene targeting in bovine fibroblasts by adeno-associated virus vectors.

Efficient PRNP gene targeting in bovine fibroblasts by adeno-associated virus vectors.
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通过腺相关病毒载体有效靶向牛成纤维细胞的 PRNP 基因。

DOI:
10.1089/15362300460743817
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发表时间:
2004
期刊:
Cloning and stem cells
影响因子:
--
通讯作者:
Russell,DavidW
Russell,DavidW
中科院分区:
--
文献类型:
--
作者:
Hirata,RoliK;Xu,Cong;Dong,Rong;Miller,DanielG;Ferguson,Stacy;Russell,DavidW

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Gene-targeted livestock can be created by combiningex vivomanipulation of cultured nuclear donor cells with cloning by nuclear transfer. However, this process can be limited by the low gene targeting frequencies obtained by transfection methods, and the limitedex vivolife span of the normal nuclear donor cells. We have developed an alternative gene targeting method based on the delivery of linear, single-stranded DNA molecules by adeno-associated virus (AAV) vectors, which can be used to introduce a variety of different mutations at single copy loci in normal human cells. Here we show that AAV vectors can efficiently target thePRNPgene encoding the prion protein PrP in bovine fetal fibroblasts, which can be used as nuclear donors to clone cattle. Cattle with bothPRNPgenes disrupted should be resistant to bovine spongiform encephalopathy.
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