Proposal of a Genome Editing System for Genetic Resistance to Tomato Spotted Wilt Virus

Viruses provoke considerable economical losses in agriculture. New molecular approaches to develop genetic resistance based on translational genomics and precision genetic modifications are highly expected. The type II Clustered, Regularly Interspaced Palindromic Repeats (CRISPR) system including Cas9 nuclease represent a promising and very powerful tool to specifically modulate the expression and activity of genes involved in biotic stress responses. In this study, we describe an approach to develop a platform system based on CRISPR system for genome editing technology in tomato. Tomato is an excellent plant for this approach considering the high-quality genome sequence, the rapid life cycle, the highly efficient in vitro plant culture and transformation protocols, Genome editing can be used to allow resistance to Tomato Spotted Wilt Virus (TSWV) infections by the successful obtainment of two specific objectives: (1) Development of a Genome Editing (GE) system using CRISPR-Cas9 system in tomato (Objective 1) and (2) test the system in inducing genetic resistance to TSWV infections. First, it will be necessary to model the molecular dynamics of key host and pathogen proteins predicting how targeted mutations affect these interactions. Then these host players will be targeted by CRISPR-Cas9 technology. The obtained plants can be evaluated for their phenotypic resistance and deeply analyzed using "omic" platforms to gain insight into gene regulatory networks of plant resistance. Outcomes of the proposed project will be essentially three: (1) Identify host target proteins interacting with pathogenic proteins and model their dynamic interactions; (2) develop a platform technology usable to obtain resistant tomatoes to TSWV inducing targeted genetic modifications in the genome; (3) facilitate the adaption of this platform to the improvement of important traits in other specialty crops.