EDGE FGT: Universal viral-based systems to accelerate gene editing research across the taxonomic range of arthropods

Pennsylvania State Univ University Park, University Park PA

Investigators

Abstract

Genetic manipulation is usually performed by injecting gene editing materials into embryos, which is difficult, expensive, and inefficient. We have identified a virus that infects the ovaries and testes of arthropods after injecting them or feeding it to them. When the virus is modified to carry gene editing materials, infected arthropods have genetically modified offspring, a technique that is much easier than traditional methods. We call this method “Germline Engineering by Viral Transduction”, or “GEValT“. GeValT is easy to use and will revolutionize gene editing for researchers working in any arthropod system. In this project, we will assess the range of arthropods that this virus infects, develop it into a gene editing delivery vehicle across multiple arthropods, develop new viral systems that act similarly for arthropods that our original virus can not infect, and disseminate information, technology and methodology to broad communities of scientists/public involved in research on animal behavior, animal physiology, insect-plant interactions, sustainable agriculture, and public health. We will host students and scientists to learn these techniques, and host two workshops to give interested scientists and students an opportunity to learn and apply these technologies to their systems of interest. All constructs/technology will be placed in public depositories to allow distribution and easy access for all interested scientists. Reagents and protocols will be shared prior to publication. The development of this biotechnology coupled with easy to use methods for arthropod genetic engineering will have broad impacts on scientific discovery, and will stimulate translational investments in the US bioeconomy industry. Genetic manipulation by embryonic microinjection is technically challenging and inefficient. We have identified a baculovirus that can infect multiple arthropod taxa where it infects diverse tissues and can deliver gene editing material to the germline, resulting in genetically modified offspring. We call this method “Germline Engineering by Viral Transduction”, or “GEValT“. GeValT is easy to use and will revolutionize gene editing methodology for researchers working in any arthropod system. Here, we will develop GeValT technologies that work across a wide range of arthropod taxa, opening the true power of this technique to all researchers interested in applying genetic engineering techniques to their questions of interest. This goal will be realized by the following four Specific Aims: 1) Characterize baculovirus infection tropism and develop compartment-specific gene transduction systems in multiple arthropods; 2) Develop GEValT methodology across multiple arthropod systems; 3) Develop novel viral infectious clone and delivery systems for arthropod taxa resistant to baculovirus infection; 4) Disseminate GEValT 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. The proposal was funded by the Enabling Discovery through GEnomics (EDGE) program. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.