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Virus-Induced Gene Silencing pp 99–107Cite as

Virus-Induced Gene Silencing Using Artificial miRNAs in Nicotiana benthamiana

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 975))

Abstract

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.

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Acknowledgments

We are very grateful to Dr. Dominique Robertson for CaLCuV vectors and to Dr. Detlef Weigel for the pRS300 vector. We also thank the National Natural Science Foundation of China (Grant nos. 31071169 and 30725002) in support of our works.

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Authors and Affiliations

  1. School of Life Sciences, Tsinghua University, Beijing, China

    Yang Tang, Yizhen Lai & Yule Liu

Authors
  1. Yang Tang
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  2. Yizhen Lai
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  3. Yule Liu
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Corresponding author

Correspondence to Yule Liu .

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Editors and Affiliations

  1. Institute of Botany, Justus-Liebig-University Gießen, Heinrich-Buff-Ring 38, Gießen, 35392, Germany

    Annette Becker

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Tang, Y., Lai, Y., Liu, Y. (2013). Virus-Induced Gene Silencing Using Artificial miRNAs in Nicotiana benthamiana . In: Becker, A. (eds) Virus-Induced Gene Silencing. Methods in Molecular Biology, vol 975. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-278-0_8

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  • DOI: https://doi.org/10.1007/978-1-62703-278-0_8

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-277-3

  • Online ISBN: 978-1-62703-278-0

  • eBook Packages: Springer Protocols

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