NSF Award
2428014
Prime editing (PE) is a recently developed, cutting-edge CRISPR/Cas gene editing technology that can accurately replace, add or remove specific pieces of DNA in an organism. Due to its high accuracy and versatility for gene editing, PE offers significant advantages over the conventional CRISPR/Cas technology and is a highly promising tool for basic plant research and applied crop breeding. This project aims to develop highly efficient and robust PE systems in rice, tomato, and poplar plants. The resulting PE tools are expected to facilitate fundamental studies in plant biology and genetic improvement of agricultural crops for important traits such as high yield, superior quality, disease resistance and abiotic stress tolerance. This project will also provide multidisciplinary research training in plant biology, functional genomics and genome engineering to postdoctoral fellows as well as graduate and undergraduate students. Outreach activities will involve local K-12 students, growers, and the general public through already established programs or newly created workshops or activities at Penn State, University of Maryland, and the National Rice Research Institute in India.<br/><br/>Prime editing is a revolutionary and advanced CRISPR/Cas genome engineering technology that enables almost all forms of precise gene editing, including base substitutions, insertions and deletions within the genome. This versatility makes PE a highly valuable tool for precise editing of plant genomes, with broad applications ranging from basic research in plant biology (e.g., epitope tagging of endogenous proteins) to practical usage in crop breeding (e.g., creating desirable alleles). Despite its successful applications in the mammalian cells, high efficiency PE in plants has only been demonstrated in rice. To fully realize the potential of PE and meet the urgent need of plant biology and crop breeding communities for precise genome editing, it is imperative to develop highly efficient and robust PE systems in plants. This project aims to optimize prime editor proteins to increase their activity and editing efficiency, improve pegRNA structure and expression for single and multiplex prime editing, and modify plant DNA repair systems to increase PE efficiency. By incorporating these innovative features and extensively testing them in rice, tomato, and poplar, highly efficient next-generation PE systems are expected to be developed for precise genome editing of both monocot and dicot plants. All data and project outcomes including plasmid vectors will be made available to the broader research communities through publications and public repositories.<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
