**AWARDS ISSUED PRIOR TO JANUARY 20, 2025, WERE FUNDED UNDER PREVIOUS ADMINISTRATIONS AND MAY NOT REFLECT THE PRIORITIES AND POLICIES OF THE CURRENT ADMINISTRATION.** GENE EDITING TECHNOLOGY PROVIDES A POWERFUL TOOL FOR SITE-SPECIFIC MANIPULATING OF DNA METHYLATION FOR CROP IMPROVEMENT, BUT ITS UNINTENDED EFFECTS ARE NOT WELL STUDIED. BETTER UNDERSTANDING OF ITS UNINTENDED EFFECTS ON DNA METHYLATION WILL ENCOURAGE ITS USE FOR DEVELOPING NEW CROPS THAT ARE MORE NUTRITIOUS AND MORE PRODUCTIVE.THIS PROJECT AIMS TO STUDY THE UNINTENDED EFFECTS OF CRISPR/DCAS-MEDIATED EDITING OF DNA METHYLATION IN TOMATO. TOMATO IS ONE OF THE MOST POPULAR AND NUTRITIOUS FOOD PRODUCTS AND ALSO ITS FRUIT QUALITY ATTRIBUTES ARE LARGELY INFLUENCED BY DNA METHYLATION EVENTS. WE WILL USE A CRISPR/DCAS-DNA METHYLTRANSFERASE SYSTEM TO PRODUCE SITE-SPECIFIC DNA METHYLATION-EDITED TOMATO PLANTS AND ALSO TO PRODUCE TOMATO PLANTS USING CONVENTIONAL TISSUE CULTURE IN-VITRO REGENERATION AND GENETIC TRANSFORMATION TECHNIQUES. TOMATO FRUITS FROM THESE PLANTS WILL BE USED FOR WHOLE GENOME BISULFITE SEQUENCING, RNA-SEQ, FRUIT QUALITY AND OTHER PHENOTYPIC ANALYSES. WE WILL CONDUCT A SIDE-BY-SIDE COMPARISON FOR THE UNINTENDED EFFECTS GENERATED FROM USING CONVENTIONAL PLANT TISSUE CULTURE, GENETIC TRANSFORMATION AND GENE EDITING TECHNOLOGIES.THE PROJECT GOAL IS CLOSELY ALIGNED WITH THE BRAG PROGRAM AREA 5F: COMPARISON OF THE TYPES AND FREQUENCIES OF NUCLEIC ACID CHANGES INTRODUCED INTO IMPORTANT CROP GENOMES, VIA GENETIC ENGINEERING TECHNIQUES VERSUS OTHER PLANT BREEDING TECHNIQUES. THE KNOWLEDGE GENERATED FROM THIS STUDY WILL AID PLANT BREEDERS PRACTICING DNA METHYLATION EDITING IN CROPS, AND FACILITATE THE POLICY- AND DECISION-MAKING PROCESS AT FEDERAL REGULATORY AGENCIES. WE WILL ALSO INCORPORATE THE SCIENCE-BASED KNOWLEDGE GENERATED FROM THIS PROJECT INTO OUR PUBLIC EDUCATION EFFORTS TO IMPROVE SOCIETY'S UNDERSTANDING ABOUT THE GENE EDITING TECHNOLOGY.

University Of Connecticut, Storrs CT

Investigators