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Improving CRISPR/Cas9 tools for precise genome editing of host plants and fungal pathogens

Yinong Yang: Pennsylvania State University

<div>The bacterial cluster regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease (Cas9) system is a powerful and versatile tool for genome editing. Using the rice-<em>Magnaporthe oryzae</em> pathosystem as a model, we have developed and improved bioinformatics and molecular tools for CRISPR/Cas9-enabled genome editing and targeted mutagenesis in the host plant and the fungal pathogen. To improve DNA editing specificity, genome-wide prediction of specific guide RNA (gRNA) spacers was performed for the rice blast fungus and eight plant species. The engineered gRNA/Cas9 system were shown to induce specific cleavages and targeted mutations at the desired genomic sites in rice and <em>M. oryzae</em>. In addition, the polycistronic tRNA-gRNA editing strategy was developed to facilitate multiplex genome editing, DNA fragment deletion, and other more sophisticated applications in plants, fungi and other organisms. With improved bioinformatics and molecular tools, the CRISPR/Cas9 system is being demonstrated to have broad applications in functional genomic analysis of host-pathogen interactions and precision breeding of crop ideotypes with enhanced disease resistance and other beneficial traits.</div>