Project Number: 5012-22000-022-24-T
Project Type: Trust Fund Cooperative Agreement
Start Date: Feb 1, 2018
End Date: Dec 31, 2020
1. Develop systems to express guide RNAs and Cas9-like enzymes from virus vector systems in crop plants. 2. Establish standard procedures to heritably modify plant genomes using viral vectors.
For these studies, infectious clones of embryo-invading plant RNA viruses will be modified to express bacterial genes and guide nucleic acids for genome editing. Because many of the bacterial genes for gene editing are larger than plant virus vectors can accommodate, the bacterial genes will be split into multiple segments and reassembled in planta using intein-mediated approaches. Guide nucleic acids will be produced from viral genomes utilizing hapten-induced or partially active ribozymes. Modified viruses will be introduced into plants (barley, corn, soybean) biolistically. Plants will be allowed to set seed, and progeny seed will be assayed for modifications to the targeted genes using high-through-put mismatch detection approaches.