Project Number: 5012-22000-023-002-A
Project Type: Cooperative Agreement
Start Date: Jul 8, 2019
End Date: Aug 14, 2022
The specific objectives of this project are: 1) Develop DNA-based methods for transient and heritable transgene expression in SCN. 2) Develop methods to artificially inoculate nematodes with positive-sense RNA viruses. 3) Construct virus-based systems to express foreign genes and silence nematode genes.
We will evaluate two approaches to achieve the goal of implementing gene drives for SCN control. First, we will modify Caenorhabditis elegans expression vectors, including vectors for expression of CRISPR-Cas9 and guide RNAs, to use SCN transcription control sequences for tissue-specific gene expression in SCN. The expression and heritability of transgenes in SCN will be assessed following particle bombardment and/or microinjection. Second, we will evaluate sexually transmitted nematode viruses to deliver gene drives to SCN germ-line cells and thus function as extra-chromosomal gene drives. Infectious clones will be prepared from positive-sense nematode viruses identified by the PIs and modified to accept foreign gene sequences. Tools developed to edit, silence and over-express nematode genes will also be used to aid in the identification and functional characterization of targets for SCN gene drives.