Location: Plant Genetics Research
Project Number: 5070-31320-001-010-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Sep 1, 2023
End Date: Aug 31, 2024
Objective 1: Develop a system to facilitate coordinated expression of multiple genes that comprise biochemical pathways. Objective 2: Evaluate biochemical impact of editing of genes involved in metabolism. Objective 3: Develop a high throughput strategy for evaluation of host factors involved in host pathways required by Influenza.
The overall approach is to investigate mechanisms that control coordinated expression. This effort requires acquisition of new knowledge, and methods to test genome edits. Three projects will be performed independently to 1) study, modify, and develop strategies for post-transcriptional coordination of independent cistrons, 2) develop and use biochemical methods to evaluate edit impacts on intermediary metabolism, and 3) develop and use methods to identify the impacts of genes and alleles on influenza replication. Specific Projects: Development of expression systems for coordinated expression of novel coding regions. Although coordinated expression of multiple coding regions is a general rule in eukaryotes, little is known of methods to harness a general mechanism. However, prokaryotes and eukaryotic viruses have developed methods to produce multiple, independent proteins from a single promoter. This project will adapt and evaluate viral and prokaryotic strategies to use single eukaryotic promoters to provide for multiple, independent proteins. Novel Intermediary Metabolism. To fully utilize genome edits developed for novel metabolic pathways, a system is required to coordinate expression of each of the individual component genes. This project will develop an expression system to allow for coordinated expression of novel genes. Host factors required for Influenza. To fully explore genome edits related to Influenza susceptibility/resistance, a high throughput strategy is required to evaluate whole genome scans and total sequence space of individual genes. This project will develop an in vitro system to rapidly identify the impact of individual genes or specific variants on Influenza replication.