Location: Soybean/maize Germplasm, Pathology, and Genetics Research
Project Number: 5012-22000-023-003-A
Project Type: Cooperative Agreement
Start Date: Jul 8, 2019
End Date: Aug 14, 2022
Objective:
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.
Approach:
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.
