Non-technical paragraph: Genetic manipulation is a powerful technique for addressing research questions in arthropods. Current approaches rely upon delivering gene-editing material to arthropod eggs by embryonic microinjection (EM). However, EM is very challenging, is limited to a small number of arthropod species, and is inefficient even in optimized species. There is a critical need to develop methods for arthropod genetic manipulation that are simple, accessible for many researchers and generally compatible for a large variety of arthropod species. We have developed a technology called Receptor-Mediated Ovary Transduction of Cargo, or ReMOT Control, to specifically deliver gene-editing cargo to the developing arthropod germline by easy injection into female arthropods during egg development. ReMOT Control can bring the power of genetic modification technology to any model or non-model species without the need for injecting embryos, allowing any lab to use these powerful tools for their research questions. Using workshops, social media, symposia, and by making reagents publically available, we will ensure that the ReMOT Control technology is available to any interested researchers. We will also use numerous outreach venues to educate the public about the benefits of these amazing tools. We will develop outreach activities and leverage existing outreach venues that engage K-12 students and members of the public to educate them about how these techniques can benefit their everyday lives.
Technical paragraph: The ReMOT Control technology is based on the identification of small peptide ligands with very specific tropism for the developing arthropod germline. These ligands are used to transduce the CRISPR/Cas9 ribonucleoprotein complex to developing eggs after injection into vitellogenic females. In this proposal, we will develop both specific and generalizable ReMOT Control technologies that work across diverse arthropod taxa, opening the true power of this technique to all researchers interested in genetic engineering techniques. ReMOT Control will break down barriers to genetic modification, allowing researchers in diverse animal systems to move beyond correlation to accurately and precisely study gene function. Our overarching conceptual goal for this proposal is nothing less than the complete democratization of gene editing capability for all researchers working in any arthropod system, be it model or non-model. This goal will be realized by the following three Specific Aims: 1) Identify species-specific and universal ligands for applying ReMOT Control technology to diverse oviparous and viviparous arthropod species; 2) Use identified ligands to transduce CRISPR/Cas9 to the host germline for targeted gene deletion and gene knock-in in diverse arthropod species; 3) Disseminate ReMOT Control information, technology and methodology to broad communities of scientists involved in research on animal behavior, animal physiology, insect-plant interactions, sustainable agriculture, and public health. By the end of the research project this transformative technology will bring genetic modification technology within the reach of everyday scientists regardless of their research system.
