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BTT EAGER: Plant genome editing and engineering via novel nanotechnology-based systems


<p>There is a national and global need to develop crop plants that yield greater food, feed, fiber and bioenergy related products. At the same time there is a need to develop crops with an increased tolerance to abiotic and biotic stress and reduced impacts on the environment. Genetic engineering and genome editing are important tools used for the study and improvement of crop plants. However, current genetic engineering/editing systems do not work efficiently, or in some cases at all in several crop species/varieties. The inability to genetically engineer certain species of plants, hinders the progress that can be made with related crops. This project is aimed at developing a new system to genetically engineer/edit plants using ultra-small, biodegradable, synthetic particles (nanoparticles). Development of such a system would enable more rapid advancement in crop genomics research and production of crops with enhanced traits and performance. In addition, the knowledge generated through this research may be utilized in the enhancement of other genome editing applications.<br />
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Genome engineering/editing systems are critical tools for the advancement of plant functional genomics and epigenomics research, and for genomics-based crop improvement efforts. Major limitations in current plant genome engineering/editing systems - such as genotype dependence, low and variable efficiencies, and/or inability to accomplish integration-free, germline editing have significantly impeded efficient, widespread utilization of these technologies for crop genomics research and genetic improvement applications. To overcome these limitations, and to increase the efficiency and breadth of plant genome editing a nanoparticle-based systems for introducing bioengineering molecular machinery into target cells capable of regenerating germline edited plants is being developed. The specific aim of the research project is: to demonstrate nanoparticle-mediated delivery of Cas9/sgRNA ribonuleoprotein (RNP) complexes into target cells in a genotype-independent manner, and successful editing of target DNA sequences in plant cells/tissues, as well as regeneration of genome-edited plants. Novel, biodegradeable nanoparticle-based systems (nanocapsule and polyplex) are used to deliver RNPs into optimal plant cell/tissue targets to generate genome-edited plant cells and tissues from which germline edited plants can be recovered. Microscopic, visual and molecular assays will be conducted to determine genome editing success, location, frequency and inheritance. What is learned here can be applied not only to plant systems, but also in non-plant editing applications. Development of an efficient, genotype-independent nanoparticle-based plant gene editing system would have significant positive impact on genome engineering-based functional genomics research and crop improvement efforts.<br />
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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.</p>

Heidi Kaeppler; Shaoqin &#039;Sarah&#039; Gong; Krishanu Saha; Xuehua Zhong
University of Wisconsin - Madison
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