Trojan Horse: Using virus-like particles as RNA delivery devices in invertebrates as a pest-control strategy

点击翻译按钮获取中文摘要

This proposal aims to spark a revolution in agricultural pest control for the 21st century. Since the middle of the twentieth century, insect pests have been controlled primarily through the use of chemical pesticides, but the use of these products is increasingly restricted because of their impact on the environment as well as animal health, including humans. However, insect pests still have the potential to cause huge amounts of damage to plant crops grown worldwide, either through direct damage caused by feeding or through the transmission of diseases to the plants. There is therefore a need to replace chemical pest control methods with safer and more specific "green" alternatives. This proposal aims to achieve this by delivering specially-designed RNA molecules to insect pests. These RNA molecules will act either through RNA interference (shutting off a gene that is critical to the insect's surival) or transient expression (causing the insect to express a protein that will be toxic). By choosing which gene sequence to target by RNA interference, or which protein to express to cause toxicity, it is possible to ensure that the product is very specific in which insect species it will affect. What is really novel about this proposal is the method that is proposed to deliver these bioactive RNA molecules to the insects: we will use a newly developed technique for packaging custom RNA molecules into protein shells that will protect the RNA as it is delivered to the insect. These protein shells are virus-like particles (VLPs) of the plant virus cowpea mosaic virus (CPMV): they are non-infectious nanoparticles that are easy to produce in large amounts in plants, and, thanks to recent discoveries, can be made to specifically package any RNA molecule of choice.During the course of this work, we will design RNA molecules for specific activity in one of five insect pest species available at the JIC Insectary. Two of these insect species are moths, two are beetles, and one is an aphid, and it is likely that delivery of bioactive RNA will work better in some insect species than others, so using a range of species increases our chance of success. We will test our bioactive RNA delivery system in these insects through feeding experiments in which we test a range of custom RNA molecules. We will also make modifications to the surface of the CPMV VLPs, by adding peptides that are intended to improve specific targeting to a particular species, or that should improve penetration of the VLPs into the insect cells. We will also compare different types of RNA molecules for RNA interference: there are different strategies that we could adopt and they may have different levels of effectiveness at causing the target insect gene to be switched off. Finally, the most promising VLP with the most promising custom RNA cargo will be tested under realistic pest control conditions to determine how good it is at being a green pesticide. Populations of the target insect will be allowed to feed on their preferred host plant in controlled conditions, and insect survival will be compared between control plants and those that have been sprayed with the VLP pesticide. Our long-term aim is to develop invertebrate-specific biocontrols to replace the reliance on chemical pesticides.The development of a novel, safe, environmentally sustainable approach to pest control in agriculture, the aim of the project, would be beneficial to all stakeholders, from the biotechnology sector to farmers and consumers.