Silencing myostatin gene by RNAi in sheep embryos

文献类型： 外文期刊

第一作者： Tang, Dayun

作者： Tang, Dayun;Zhu, Huabin;Zhang, Yan;Zhao, Xueming;Chen, Xiaoliang;Du, Weihua;Wang, Dong;Lin, Xiukun;Tang, Dayun;Chen, Hanzhong;Chen, Xiaoliang;Tang, Dayun;Wu, Jianmin

作者机构：

关键词： RNAi;Lentivirus;Transgenic Embryo;Sheep

期刊名称：JOURNAL OF BIOTECHNOLOGY （ 影响因子：3.307； 五年影响因子：3.778 ）

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收录情况： SCI

摘要： Myostatin (MSTN) gene is described as a negative regulator of the skeletal muscle growth. Controlling MSTN gene expression by genetic manipulation could accelerate the muscle growth and meat production of livestock animals. In the present study, several siRNAs targeting sheep MSTN gene were designed and their interfering efficiency was evaluated in vitro. The present study showed that one of the siRNAs, PSL1, could down-regulate the expression of MSTN significantly. PSL1 was ligated into lentivirus vector, GP-Supersilencing, to construct a siRNA expression lentivirus vector. Fibroblast cells were infected by lentivirus particles and positive cells were isolated by flow cytometry. Nucleus of the positive cell was transferred into enucleated oocytes of sheep. The present study showed that 99.4% of the sorted cells displayed green fluorescence. After enucleation of oocytes with microinjection, about 20% of reconstructed embryos can be developed into morulas, and strong green fluorescence could be observed using a fluorescence microscope. This method can be available to produce transgenic cell line for somatic cell nucleus transfer for transgenic animals

分类号： Q81

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