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1 results

  • Effect of using green fluorescent protein double-stranded RNA as non-target negative control in Nasonia vitripennis RNA interference assays

  • Julien Rougeot, Yidong Wang, Eveline C. Verhulst
  • Journal:
    Experimental Results / Volume 2 / 2021
    Published online by Cambridge University Press:
    10 February 2021, e11
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  • RNA interference (RNAi) is a technique used in many insects to study gene function. However, prior research suggests possible off-target effects when using Green Fluorescent Protein (GFP) sequence as a non-target control. We used a transcriptomic approach to study the effect of GFP RNAi (GFP-i) in Nasonia vitripennis, a widely used parasitoid wasp model system. Our study identified 3.4% of total genes being differentially expressed in response to GFP-i. A subset of these genes appears involved in microtubule and sperm functions. In silico analysis identified 17 potential off-targets, of which only one was differentially expressed after GFP-i. We suggest the primary cause for differential expression after GFP-i is the non-specific activation of the RNAi machinery at the injection site, and a potentially disturbed spermatogenesis. Still, we advise that any RNAi study involving the genes deregulated in this study, exercises caution in drawing conclusions and uses a different non-target control.