RNAi gene knockdown in the poultry red mite, Dermanyssus gallinae (De Geer 1778), a tool for functional genomics

Wan Chen, Kathryn Bartley, Francesca Nunn, Alan Bowman, Jeremy Sternberg, Stewart T G Burgess, Alasdair J Nisbet, Daniel R G Price*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)
5 Downloads (Pure)

Abstract

Background
The avian haematophagous ectoparasite Dermanyssus gallinae, commonly known as the poultry red mite, causes significant economic losses to the egg-laying industry worldwide and also represents a significant welfare threat. Current acaricide-based controls are unsustainable due to the mite’s ability to rapidly develop resistance, thus developing a novel sustainable means of control for D. gallinae is a priority. RNA interference (RNAi)-mediated gene silencing is a valuable tool for studying gene function in non-model organisms, but is also emerging as a novel tool for parasite control.

Methods
Here we use an in silico approach to identify core RNAi pathway genes in the recently sequenced D. gallinae genome. In addition we utilise an in vitro feeding device to deliver double-stranded (ds) RNA to D. gallinae targeting the D. gallinae vATPase subunit A (Dg vATPase A) gene and monitor gene knockdown using quantitative PCR (qPCR).

Results
Core components of the small interfering RNA (siRNA) and microRNA (miRNA) pathways were identified in D. gallinae, which indicates that these gene silencing pathways are likely functional. Strikingly, the P-element-induced wimpy testis (PIWI)-interacting RNA (piRNA) pathway was absent in D. gallinae. In addition, feeding Dg vATPase A dsRNA to adult female D. gallinae resulted in silencing of the targeted gene compared to control mites fed non-specific lacZ dsRNA. In D. gallinae, dsRNA-mediated gene knockdown was rapid, being detectable 24 h after oral delivery of the dsRNA, and persisted for at least 120 h.

Conclusions
This study shows the presence of core RNAi machinery components in the D. gallinae genome. In addition, we have developed a robust RNAi methodology for targeting genes in D. gallinae that will be of value for studying genes of unknown function and validating potential control targets in D. gallinae.
Original languageEnglish
Article number57
Number of pages13
JournalParasites & Vectors
Volume14
Issue number1
DOIs
Publication statusPublished - 18 Jan 2021

Bibliographical note

The authors gratefully acknowledge funding for this project from the Scottish Government Rural Affairs, Food and the Environment (RAFE) Strategic Research Portfolio 2016-2021. DRGP is supported by a research fellowship provided by the Moredun Foundation. WC is supported by a studentship provided by the University of Aberdeen and the Moredun Foundation.

Keywords

  • RNA Interference
  • Poultry red mite
  • Functional genomics
  • Gene Silencing
  • Gene knockdown
  • Gene silencing
  • RNA interference

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