TOOLS FOR RAPID GENETIC TRANSFORMATION OF BEES

Pollinator populations are in decline globally, and existing toolkits of management practices have been inadequate for halting and reversing losses. Recently developed genomic editing technologies can be applied to study the mechanistic basis of diverse stressors on pollinators and develop novel solutions to improve their ability to withstand multiple stressors in the field. Current genomic editing technologies rely upon delivering DNA or endonucleases to preblastoderm embryos via embryonic microinjection. Embryonic microinjection is technically challenging, limited to a small number of arthropod taxa, and is inefficient even in optimized species. A critical need exists to develop methods for arthropod genetic manipulation that are simple, accessible, and compatible for a large variety of arthropod species (including pollinators). During oogenesis, arthropods transfer yolk protein precursors (such as vitellogenin) to developing oocytes by receptor-mediated endocytosis (RME). We have developed a technique (Receptor-Mediated Ovary Transduction of Cargo; ReMOT Control), which exploits RME to transduce cargo into the developing germline for heritable editing of the chromosomal sequence. We will use ReMOT Control to develop easy and efficient genetic manipulation technologies for the native pollinator Bombus impatiens. In Aim One, we will assess the ability for B. impatiens vitellogenin (BiVg) to transduce protein cargo (GFP) into bee oocytes. In Aim Two, we will use BiVg to transduce into the germline transcription activator-like effector nucleases (TALENs) targeting an eye color gene for heritable editing of the genetic sequence. Our experiments will revolutionize studies of managed bee pollinators, and can be applied to other economically important Hymenoptera.