Tropical super flies: Integrating Cas9 into Drosophila ananassae and its phenotypic effects

Ectotherms such as insects are animals whose body temperature largely depends on ambient temperature and temperature variations provide a selection pressure affecting the geographical distribution of these species. However, over the course of evolution, some insect species managed to colonize environments characterized by various temperature ranges. Therefore, insects provide an excellent study system to investigate the basis of adaptation to temperature changes and extremes. We are generally using the vinegar fly Drosophila ananassae as a model system to investigate the genetic basis of cold tolerance. This species has expanded from its tropical ancestral range to more temperate regions resulting in a cosmopolitan, domestic distribution. Previously, we identified candidate genes significantly associated with cold tolerance in this species. We now established molecular genetic tools to assess the function of these genes. Using CRISPR/Cas9 methodology for genome editing and the PiggyBac system, the Cas9 enzyme was successfully integrated into the genome of three fly strains with different levels of cold tolerance. We further report on preliminary findings that the Cas9 integration itself did not have a consistent effect on tolerance to cold. In conclusion, we offer with our study the molecular tools that allow studying stress-related candidate genes in D. ananassae in the future. In addition, we point out and provide guidance on the challenges that come with genome editing in a non-model species.

Automated summary

Ectotherms, such as insects, have body temperatures that depend on ambient temperature, and temperature variations affect the distribution of these species. Insects provide a good study system for investigating adaptation to temperature changes, and the vinegar fly Drosophila ananassae is being used as a model to study the genetic basis of cold tolerance. Molecular genetic tools, such as CRISPR/Cas9 and the PiggyBac system, have been established to assess the function of candidate genes associated with cold tolerance. The integration of Cas9 enzyme did not consistently affect cold tolerance in this species.