CAREER: Elucidating trans-kingdom horizontal gene transfer mechanisms to improve plant genetic engineering

Stanford University, Stanford CA

Investigators

Abstract

Plants lie at the heart of many potential solutions to the climate crisis. Yet, it can take years to develop plants with new, useful features. Plant transformation and negative public opinion are widely recognized as the main bottlenecks to engineering new plant varieties. This team will pursue an integrated research and education plan to spur innovation in plant genetic engineering. The research will generate foundational knowledge related trans-kingdom DNA transfer between Agrobacterium and plants, which will be used to develop plant transformation tools that improve the speed and complexity of genetic manipulation achievable in plants. In parallel, the PI will develop a summer program for students enrolled in their National Education Equity Lab course: BIOE80 “Introduction to Bioengineering.” The PI already works with the National Education Equity lab – a non-profit that provides low-income (Title 1) high school students with an opportunity to earn free college credits by taking college courses from college professors. The proposed summer addition to BIOE80 will give students an opportunity to gain hands-on experience with the genetic engineering techniques learned in their lecture-based course. It will serve as much needed bridge between the students’ theoretical understanding of bioengineering concepts and the research experience needed to pursue internships and higher education in STEM field. Ultimately, the course and proposed research should strengthen U.S. plant engineering discourse and research should help more fully realize the potential of plant biotechnology for a sustainable future. Trans-kingdom transfer of DNA from Agrobacterium to plants is a stunning biological feat and the basis of powerful plant biotechnology tools. Despite being the most commonly used tool for introducing new DNA to plants, several important gaps in our understanding of Agrobacterium-mediated DNA transfer remain – including the dynamics and mechanisms of transferred DNA (T-DNA) integration into the plant genome. This knowledge gap has prevented the generation of plant genome engineering tools that can be used to reliably control the insertion location of transgenes in plants’ genomes and limits the speed and complexity of genetic perturbation achievable in plants. The proposed research will investigate the temporal dynamics of T-DNA delivery and T-DNA fate (integration or degradation) in plant cells after the two most common plant transformation procedures: floral dip and in vitro conjugation to callus. The team will use a new synthetic genetic system, developed by the PI, that can record T-DNA delivery to dissect these complex biological processes with unprecedented precision. Then, newly acquired information will be used to create molecular tools for plant genome editing without T-DNA integration. This work will contribute to the bioeconomy by generating the foundational knowledge needed to develop increasingly sophisticated plant genome engineering tools and setting the stage for the long-term goal of precisely engineering plants in order to improve climate resilience. 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.