Targeted microRNA expression in dairy cattle directs production of β-lactoglobulin-free, high-casein milk
Targeted microRNA expression in dairy cattle directs production of β-lactoglobulin-free, high-casein milk
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DOI：
10.1073/pnas.1210057109
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发表时间：
2012-10-16
影响因子：
11.1
通讯作者：
Laible, Goetz
中科院分区：
文献类型：
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作者：
Jabed, Anower;Wagner, Stefan;Laible, Goetz
来自奶牛的牛奶含有蛋白质-乳球蛋白（BLG），这是不存在于人类的牛奶。由于它是一种主要的牛奶过敏原，我们希望通过RNAi降低牛奶中的BLG水平。在体外筛选10个microrna (mirna)，无论是单独的还是串联的组合，确定了几个实现高达98%的BLG敲除。在表达羊BLG的小鼠乳腺模型中表达一个串联结构，使羊乳中BLG的表达降低96%。在体内验证之后，我们生产了一只转基因小牛，设计表达这些串联mirna。对这头小牛的激素诱导乳的分析表明，BLG的缺失和所有酪蛋白乳蛋白的同时增加。研究结果证实了miRNA介导的牛致敏乳蛋白的消耗，并验证了靶向miRNA表达是改变牛奶成分和其他牲畜性状的有效策略。
Milk from dairy cows contains the protein beta-lactoglobulin (BLG), which is not present in human milk. As it is a major milk allergen, we wished to decrease BLG levels in milk by RNAi. In vitro screening of 10 microRNAs (miRNAs), either individually or in tandem combinations, identified several that achieved as much as a 98% knockdown of BLG. One tandem construct was expressed in the mammary gland of an ovine BLG-expressing mouse model, resulting in 96% knockdown of ovine BLG in milk. Following this in vivo validation, we produced a transgenic calf, engineered to express these tandem miRNAs. Analysis of hormonally induced milk from this calf demonstrated absence of BLG and a concurrent increase of all casein milk proteins. The findings demonstrate miRNA-mediated depletion of an allergenic milk protein in cattle and validate targeted miRNA expression as an effective strategy to alter milk composition and other livestock traits.