PlantSynBio: A Novel CRISPR SynBio Tool for Investigating and Reprogramming the Regulation of Alkaloid Biosynthesis in Catharanthus roseus

点击翻译按钮获取中文摘要

Plants have developed elaborate mechanisms for responding rapidly and specifically to changes in their environment. For instance, in response to attack by pathogens, insects, or herbivores, plants strategically redirect their energy and resources from growth and development towards the production of defense-related compounds. The production of defense-related compounds is also carefully controlled by the plant to limit the costly expenditure of energy and resources. The fundamental scientific contribution of this research is to understand how plants rapidly and efficiently coordinate and control this defense response at the molecular level. This project develops and implements novel experimental tools to characterize and investigate the processes that regulate the production of terpenoid indole alkaloids in the plant Catharanthus roseus. The study has the potential to hasten the investigation and consequently the solution to challenges in engineering hardy crops and producing important medicines while providing fundamental understanding of plants. There is broad interdisciplinary dissemination of this science through teaching and the mentoring of high school, undergraduate, and graduate students.To understand how plants rapidly and efficiently coordinate and control their defense response at the molecular level, the model system selected for this research is the Catharanthus roseus plant. This plant produces the valuable compounds known as terpenoid indole alkaloids (TIAs) upon herbivory; important TIAs from this plant include the chemotherapeutic drugs, vinblastine and vincristine. To elucidate the regulation of TIA biosynthesis, the first aim of this research identifies potential transcription factors involved in controlling TIA biosynthesis. The second aim designs, builds, and tests a novel synthetic biology tool based on CRISPR-dCas, to simultaneously but independently regulate the expression of multiple transcription factors. This validated synthetic biology tool is then applied to reprogram the regulation and production of TIAs in transgenic plants or tissue cultures. This research contributes a novel synthetic biology tool that will be made available for diverse applications in the plant community, will reveal how plants rapidly and efficiently coordinate and control the defense response, and will enable strategies for reprogramming and enhancing TIA production.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.