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慢病毒介导的 RNAi 稳定沉默山羊胎儿成纤维细胞中的β-乳球蛋白(BLG)基因。

Stable silencing of β-lactoglobulin (BLG) gene by lentivirus-mediated RNAi in goat fetal fibroblasts.

机构信息

College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, People's Republic of China.

出版信息

Genet Mol Biol. 2012 Jul;35(3):680-5. doi: 10.1590/S1415-47572012005000042. Epub 2012 Jul 5.

Abstract

β-lactoglobulin (BLG), a dominant allergen in goat milk, is difficult to remove by traditional biochemical methods. Its elimination from goat milk by genetic modification therefore poses a major challenge for modern goat breeders. A shRNA targeting BLG mRNA with high interference efficiency was identified, with which lentiviral vectors were used for mediating stable shRNA interference in goat-fetal fibroblast cells. Apart from high efficiency in the knockdown of BLG expression in these cells, lentivector-mediated RNAi manifested stable integration into the goat genome itself. Consequently, an in vitro model for goat BLG-content control was compiled, and a goat-cell line for accompanying transgenetic goat production created.

摘要

β-乳球蛋白(BLG)是羊奶中的主要过敏原,用传统的生化方法很难去除。因此,通过基因修饰从羊奶中去除它对现代奶山羊饲养者来说是一个重大挑战。我们筛选出了一种靶向 BLG mRNA 的高效干扰 shRNA,并用慢病毒载体介导其在山羊胎儿成纤维细胞中稳定的 shRNA 干扰。除了在这些细胞中高效敲低 BLG 表达外,慢病毒介导的 RNAi 还能稳定整合到山羊基因组本身。因此,我们构建了体外控制羊奶 BLG 含量的模型,并创建了伴随转基因山羊生产的山羊细胞系。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716a/3459420/71f2d0f9b6d7/gmb-35-3-680-gfig1.jpg

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