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cDNA Libraries for Virus-Induced Gene Silencing

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Plant Epigenetics

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 631))

Abstract

Virus-induced gene silencing (VIGS) exploits endogenous plant antiviral defense mechanisms to posttranscriptionally silence the expression of targeted plant genes. VIGS is quick and relatively easy to perform and therefore serves as a powerful tool for high-throughput functional genomics in plants. Combined with the use of subtractive cDNA libraries for generating a collection of VIGS-ready cDNA inserts, VIGS can be utilized to screen a large number of genes to determine phenotypes resulting from the knockdown/knockout of gene function. Taking into account the optimal insert design for VIGS, we describe a methodology for producing VIGS-ready cDNA libraries enriched for inserts relevant to the biological process of interest.

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Acknowledgments

We are grateful to S. Dinesh-Kumar (Yale University) for generously providing the TRV vectors. We also thank the DNA Service Unit at NRC-PBI for EST sequencing, Jacek Nowak and Kannan Vijayan for bioinformatic analysis, and Sandra Polvi for assistance in plant cultivation. This is manuscript NRCC #50134.

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Authors and Affiliations

  1. NRC Plant Biotechnology Institute, Saskatoon, SK, Canada

    Andrea T. Todd, Enwu Liu & Jonathan E. Page

Authors
  1. Andrea T. Todd
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  2. Enwu Liu
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  3. Jonathan E. Page
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Editor information

Editors and Affiliations

  1. Dept. Biological Sciences, University of Lethbridge, University Drive 4401, Lethbridge, T1K 3M4, Canada

    Igor Kovalchuk

  2. Dept. Biological Sciences, University of Lethbridge, University Drive 4401, Lethbridge, T1K 3M4, Canada

    Franz J. Zemp

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Todd, A.T., Liu, E., Page, J.E. (2010). cDNA Libraries for Virus-Induced Gene Silencing. In: Kovalchuk, I., Zemp, F. (eds) Plant Epigenetics. Methods in Molecular Biology™, vol 631. Humana Press. https://doi.org/10.1007/978-1-60761-646-7_16

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  • DOI: https://doi.org/10.1007/978-1-60761-646-7_16

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-645-0

  • Online ISBN: 978-1-60761-646-7

  • eBook Packages: Springer Protocols

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