I-Corps: Translation potential of DNA-free gene editing technology for crop improvement

This I-Corps project is based on the development of the commercial potential of a DNA-free crop gene-editing platform. Traditional breeding methods and existing gene-editing platforms are slow, costly, and do not work for all crops. This technology bypasses traditional barriers by providing a precise, tissue-culture-free, DNA-free gene editing method that works across any flowering plant or variety. The goal is to cut the traditional 10-year crop breeding cycle in half. This technology may shorten breeding cycles and cuts costs, as well as reduce unintended genetic changes. In addition, it is a genetically modified organism (GMO)-free platform and may provide a new plant breeding tool, to improve crop production and safeguard the future food supply. Users may benefit from faster research pipelines, and improved consistency in outcomes compared with traditional tissue culture. For the industry, this means faster product development and a competitive advantage. For academia, it enables access to high-quality gene-edited lines to achieve higher research productivity. This I-Corps project utilizes experiential learning coupled with first-hand investigation of the industry ecosystem to assess the translation potential of a DNA- and tissue culture-free, genotype- and species-independent gene editing technology for crops. Traditional genetic engineering remains constrained by tissue-culture recalcitrance, species- and genotype-dependence, and lengthy, expensive breeding cycles. To overcome these critical challenges, the technology uses a tissue culture-free, species- and genotype-independent transformation approach using pollen as the primary delivery vessel for pre-assembled Cas-gRNA ribonucleoprotein (RNP) complexes used in CRISPR systems for gene editing. Synthesized Cas endonucleases are combined in vitro with guide RNA to form fully functional RNPs, which are directly delivered into mature pollen cells. Upon delivery, the active RNP edits the target genome without recombinant plasmid integration, foreign DNA introduction, or vector insertion. Subsequent fertilization with egg cells generates non-genetically modified organism (GMO) mutant progeny in the first generation, bypassing tissue-culture regeneration. Previous research demonstrated efficient target genome modification and robust editing frequencies in pollen-derived seeds across diverse flowering crop species. This technology may provide plant breeders and commercial agricultural researchers with an efficient, cost-effective tool to accelerate trait development, minimize regulatory burdens, shorten crop breeding timelines, and cut costs of crop improvement. 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.