Sonoporation-Mediated Delivery of Reagents into Insect Embryos

Project description: Sonoporation is a technique that uses ultrasound in conjunction with lipid microbubbles to permeabilize membranes and deliver various molecules into cells. Our lab is pioneering the use of this technique in insect embryos, including a pilot project funded by CAMTech to deliver CRISPR/Cas9 reagents to Southern green stink bugs (SGSB). Sonoporation is low-cost (~$1,000 in equipment) and high-throughput (batches of eggs can be processed in minutes) and is thus an attractive option for embryonic delivery in insects that are recalcitrant to microinjection. Here, we will expand this approach in SGSB and other insect pests and test the extent to which sonoporation can become a flexible, all-in-one tool for delivering reagents into insect embryos. Applications include but are not limited to delivery of CRISPR/Cas9 and other transformation reagents, delivery of small molecules and novel chemistries that could function as potential ovicides, and introduction of compatible solutes for cryopreservation. Our project has two specific objectives: 1) Expand sonoporation capabilities in SGSB to include transgenic overexpression, RNAi, and delivery of small molecules; 2) Develop sonoporation methods for diverse insect pests of interest to industry and the entomological research community. Together, this work will create a low cost and technically accessible solution for delivering reagents into insect embryos.