A simplified method for constructing artificial microRNAs based on the osa-MIR528 precursor.
作者： Fei Yan , Yuwen Lu , Gentu Wu , Jiejun Peng , Hongying Zheng
DOI: 10.1016/J.JBIOTEC.2012.02.015
关键词: microRNA 、 Computational biology 、 Gene 、 Nicotiana benthamiana 、 Biology 、 Gene silencing 、 Genetics
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 construct amiRNAs, requires four PCRs. We describe a simplified constructing amiRNA based on the osa-MIR528 backbone using one PCR step by addition prolonging sequence primers. length needed precursor was determined. Using this method, we constructed targeting Nicotiana benthamiana UPF1 gene and showed that it functioned silencing expression leaves when expressed transiently.
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