A simplified method for constructing artificial microRNAs based on the osa-MIR528 precursor

文献类型: 外文期刊

第一作者: Yan, Fei

作者: Yan, Fei;Lu, Yuwen;Wu, Gentu;Peng, Jiejun;Zheng, Hongying;Lin, Lin;Chen, Jianping;Wu, Gentu;Peng, Jiejun

作者机构:

关键词: Artificial microRNA;Construction;MicroRNA;Osa-MIR528

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: Artificial miRNAs (amiRNAs) have been used successfully in various plants to silence endogenous genes or viral RNAs. The method of Schwab et al., widely used to construct amiRNAs, requires four PCRs. We describe a simplified method of constructing amiRNA based on the osa-MIR528 backbone using one PCR step by addition of prolonging sequence to the primers. The length of prolonging sequence needed in the osa-MIR528 precursor was determined. Using this method, we constructed amiRNA targeting the Nicotiana benthamiana UPF1 gene and showed that it functioned in silencing UPF1 expression in leaves when expressed transiently

分类号: Q81

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