Direct Transfer of Synthetic Double-Stranded RNA into Protoplasts of Arabidopsis thaliana

  • Ha-il Jung
  • Zhiyang Zhai
  • Olena K. Vatamaniuk
Part of the Methods in Molecular Biology book series (MIMB, volume 744)


Double-stranded (ds) RNA interference (RNAi) is widely used as a reverse genetic approach for functional analysis of plant genes. Constitutive or transient RNAi effects in plants have been achieved via generating stable transformants expressing dsRNAs or artificial microRNAs (amiRNAs) in planta or by viral-induced gene silencing (VIGS). Although these tools provide outstanding resources for functional genomics, they require generation of vectors expressing dsRNAs or amiRNAs against targeted genes, transformation and propagation of transformed plants, or maintenance of multiple VIGS lines and thus impose time, labor, and space requirements. As we showed recently, these limitations can be circumvented by inducing RNAi effects in protoplasts via transfecting them with in vitro-synthesized dsRNAs. In this chapter we detail the procedure for transient gene silencing in protoplasts using synthetic dsRNAs and provide examples of approaches for subsequent functional analyses.

Key words

In vitro-synthesized dsRNA protoplasts protoplast viability assays RNAi 


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Ha-il Jung
    • 1
  • Zhiyang Zhai
    • 1
  • Olena K. Vatamaniuk
    • 1
  1. 1.Department of Crop and Soil SciencesCornell UniversityIthacaUSA

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