Effect of using green fluorescent protein double-stranded RNA as non-target negative control in Nasonia vitripennis RNA interference assays
| Title: | Effect of using green fluorescent protein double-stranded RNA as non-target negative control in Nasonia vitripennis RNA interference assays |
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| Authors: | Julien Rougeot, Yidong Wang, Eveline C. Verhulst, Michael Nevels |
| Source: | Experimental Results, Vol 2 (2021) |
| Publisher Information: | Cambridge University Press, 2021. |
| Publication Year: | 2021 |
| Collection: | LCC:Technology LCC:Medicine LCC:Science |
| Subject Terms: | RNA interference, Nasonia, RNA sequencing, Technology, Medicine, Science |
| More Details: | 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. |
| Document Type: | article |
| File Description: | electronic resource |
| Language: | English |
| ISSN: | 2516-712X |
| Relation: | https://www.cambridge.org/core/product/identifier/S2516712X20000672/type/journal_article; https://doaj.org/toc/2516-712X |
| DOI: | 10.1017/exp.2020.67 |
| Access URL: | https://doaj.org/article/15ce67dcd3dc4df7ab96327161d3ec7a |
| Accession Number: | edsdoj.15ce67dcd3dc4df7ab96327161d3ec7a |
| Database: | Directory of Open Access Journals |
| ISSN: | 2516712X |
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| DOI: | 10.1017/exp.2020.67 |
| Published in: | Experimental Results |
| Language: | English |