Construction of Mismatched Inverted Repeat (IR) Silencing Vectors for Maximizing IR Stability and Effective Gene Silencing in Plants

  • M. E. Chrissie Rey
  • Johan Harmse
  • Sarah H. Taylor
  • Patrick Arbuthnot
  • Marc S. Weinberg
Part of the Methods in Molecular Biology book series (MIMB, volume 1287)


Inverted repeat (IR) RNA silencing vectors containing homologous fragments of target endogenous plant genes, or pathogen genes, are the most widely used vectors to either study the function of genes involved in biotic stress or silence pathogens to induce plant resistance, respectively. RNA silencing has been exploited to produce transgenic plants with resistance to viral pathogens via posttranscriptional gene silencing (PTGS). In some cases, this technology is difficult to apply due to the instability of IR constructs during cloning and plant transformation. We have therefore developed a robust method for the production of long IR vector constructs by introducing base pair mismatches in the form of cytosine to thymine mutations on the sense arm by exposure to sodium bisulfite prior to assembly of the IR.

Key words

Inverted repeat RNA silencing Plants Mismatches Sodium bisulfite 



We would like to thank the South African National Research Foundation, Casquip Starch Manufacturing Pty Ltd. (Jim Casey), and The Technical Innovation Agency for financial contributions to this project.


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • M. E. Chrissie Rey
    • 1
  • Johan Harmse
    • 1
  • Sarah H. Taylor
    • 1
  • Patrick Arbuthnot
    • 1
  • Marc S. Weinberg
    • 1
  1. 1.School of Molecular and Cell BiologyUniversity of the WitwatersrandJohannesburgSouth Africa

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