Single cell RNA-seq profiling of ovaries after dnmt1 gene expression knockdown

As insects continue to evolve resistance to commercially available pesticides, the hunt for novel candidate genes and pathways to target as part of integrated pest management strategies gains more importance. RNA interference (RNAi) technologies, which reduces the gene expression of a targeted gene, are becoming newer additions to existing management strategies of agricultural pests. A prerequisite for RNAi technologies successful deployment is identifying candidate genes of known effect and mechanism to bias against non-target consequences. We have been investigating the mechanism of action of DNA methyltranferase 1 (dnmt1) in insect reproduction. When dnmt1 gene expression is reduced with RNAi, both the number of eggs produced and the viability of embryos that are produced is severely reduced. We are now using single cell RNA-sequencing to help us to understand the cell population impacted, the genetic network that dnmt1 is embedded within, and some of the genetic perturbations that are caused from reducing its expression. This work provides a foundation to assess the viability of using dnmt1 with RNAi technologies as part of pest management strategies within Hemiptera in the future. Although our work is targeted at finding candidate genes for pest management strategies, it will also provide a greater basic understanding of insect ovaries outside of existing model species, most notably Drosophila melanogaster.