Virus-induced gene silencing using artificial miRNAs in Nicotiana benthamiana.

Virus-induced gene silencing using artificial microRNAs (MIR VIGS) is a newly developed technique for plant reverse genetic studies. Traditional virus-induced gene silencing (VIGS) assays introduce a large gene fragment, which is expressed and then converted into small RNAs by the endogenous siRNA-based gene silencing machinery of the plant host. By contrast, MIR VIGS uses well-designed miRNAs to induce RNA-mediated silencing of the target gene. Using a single artificial miRNA can provide greater specificity by reducing off-target effects. Here, we describe a detailed protocol for MIR VIGS in Nicotiana benthamiana using a modified Cabbage leaf curl virus (CaLCuV)-based vector.

Citation

Yang Tang, Yizhen Lai, Yule Liu. Virus-induced gene silencing using artificial miRNAs in Nicotiana benthamiana. Methods in molecular biology (Clifton, N.J.). 2013;975:99-107

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PMID: 23386298

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