Using CRISPR/Cas9 genome modification to understand the genetic basis of insecticide resistance: Drosophila and beyond

Chemical insecticides are a major tool for the control of many of the world's most damaging arthropod pests. However, their intensive application is often associated with the emergence of resistance, sometimes with serious implications for sustainable pest control. To mitigate failure of insecticide-based control tools, the mechanisms by which insects have evolved resistance must be elucidated. This includes both identification and functional characterization of putative resistance genes and/or mutations. Research on this topic has been greatly facilitated by using powerful genetic model insects like Drosophila melanogaster, and more recently by advances in genome modification technology, notably CRISPR/Cas9. Here, we present the advances that have been made through the application of genome modification technology in insecticide resistance research. The majority of the work conducted in the field to date has made use of genetic tools and resources available in D. melanogaster. This has greatly enhanced our understanding of resistance mechanisms, especially those mediated by insensitivity of the pesticide target-site. We discuss this progress for a series of different insecticide targets, but also report a number of unsuccessful or inconclusive attempts that highlight some inherent limitations of using Drosophila to characterize resistance mechanisms identified in arthropod pests. We also discuss an experimental framework that may circumvent current limitations while retaining the genetic versatility and robustness that Drosophila has to offer. Finally, we describe examples of direct CRISPR/Cas9 use in non-model pest species, an approach that will likely find much wider application in the near future. [Display omitted] •CRISPR/Cas9 genome modification is a powerful tool to study insecticide resistance.•Genome modified Drosophila has a growing use in resistance studies, but also inherent limitations.•Certain limitations may be overcome by applying CRISPR/Cas9 in pest species.