Development of efficacious ribonucleic acid-based formulations for the control of gypsy moth

Recipient Organization
NANOSUR LLC
1951 NW 7TH AVE
MIAMI,FL 331361104

Performing Department
(N/A)

Non Technical Summary
The gypsy moth is a very difficult-to-control insect pest that has a significant impact on many economic sectors, including forestry. Current biological insecticides used in gypsy moth control have not been efficacious enough and chemical insecticides are not selective and cause unnecessary ecological damage. The ranking of the gypsy moth as the third most expensive invasive insect in the world, with an estimated annual 3.2 billion dollars cost just in North America is indicative of the impact of this pest. Ribonucleic acid interference (RNAi) represents a viable option for the selective control of the gypsy moth. However, current RNAi triggers are quantitatively inefficient, in part due to their instability and deficient transport properties across biological membranes. NanoSUR is proposing the development of RNAi triggers efficacious against thegypsy moth with enhanced stability and delivery properties.This Project supports theUSDA strategic plan goal ofensuring theproductive and sustainable use of our national forest system lands. This Project specifically contributes to the economic health of rural communities through the use of and access to availableopportunities and ensures that lands and watersheds are sustainable, healthy and productive. This Projectalso contributes tothe reduction of the adverse impacts from insects, diseases, and wildfires.

Animal Health Component

20%

Research Effort Categories

Basic

10%

Applied

20%

Developmental

70%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
123	0699	1060	50%
124	0530	1060	50%

Knowledge Area
123 - Management and Sustainability of Forest Resources; 124 - Urban Forestry;

Subject Of Investigation
0530 - Parks and urban green space; 0699 - Trees, forests, and forest products, general;

Field Of Science
1060 - Biology (whole systems);

Keywords

Goals / Objectives
Gypsy moth (GM, Lymantria dispar) is an invasive pest with significant socioeconomic impact. The goal of this project is to enable a commercially viable and environmentally safe method to control GM.The following are the objectives that will be undertaken to achieve the goal aboveObtain an improved RNAi trigger by evaluating different RNA-modifying reagents using an artificial diet assay. The readouts of this assay will be larva mortality counts and gene expression levels (by qPCR).Evaluatethe efficacy of the most promising candidate identified in the previous objective to control the GM pest larvae in a leaf disk assay.

Project Methods
The following describes the methods that will be used to conduct the project and the evaluation of the results obtained:Synthesis and testing of the modified double stranded ribonucleic acid (MdsRNA) materials preparedand using artificial diet and qPCR assays.Scale upand testing of the best preparation advanced fromthe previous stepusing a leaf disk assay.

Progress 07/01/21 to 08/31/22

Outputs
Target Audience:This application addresses Research Topic 8.1 (Forests and Related Resources) FY21 Research Priorities 1 Growth and Yield. This topic supports FY2018-2022 USDA Strategic Plan Goal 6: Ensure Productive and Sustainable Use of Our National Forest System Lands. This application specifically addresses SUB-GOALS 6.1 and 6.2: respectively "Contribute to the Economic Health of Rural Communities Through Use and Access Opportunities" and "Ensure Lands and Watersheds Are Sustainable, Healthy, and Productive" and contributes to the Strategy: " ... to more effectively address forest health conditions ... " and to the Outcome: " ... USDA protects water quality; reduces the adverse impacts from insects, diseases, and wildfires...". Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The overall goal of this proposal was achieved. Together with our ARS collaborators, we identified a NanoSUR's chemically modified dsRNA (J1-6) targeting a gene essential to the survival of the invasive Lepidopteran pest European gypsy moth (EGM, Lymantria dispar dispar L.), which showed significant efficacy against EGM larvae in in vivo efficacy models. Using our proprietary knowledge from the identification and development of potent modified dsRNA triggers for two other Lepidopteran global pests, namely Diamondback moth (Plutella xyllostella) and Fall armyworm (Spodoptera frugiperda) we selected 3 modifying groups and 2 EGM sequences, dsSEQ1 and dsSEQ2, to produce NanoSUR's modified dsRNA (MdsRNA). These MdsRNAs were tested in artificial diet (AD) and leaf disc assays with 3rd instar L. dispar larvae and qPCR was used to measure gene expression levels. Three of the tested MdsRNAs, J1-6 (M4dsSEQ1), J1-4 (M2dsSEQ1), and J2-2 (M2dsSEQ2) produced the biggest depression in gene expressions and have been advanced to further studies.

Publications