**AWARDS ISSUED PRIOR TO JANUARY 20, 2025, WERE FUNDED UNDER PREVIOUS ADMINISTRATIONS AND MAY NOT REFLECT THE PRIORITIES AND POLICIES OF THE CURRENT ADMINISTRATION.** MICROBES CAN BE ENGINEERED TO PERFORM NEW FUNCTIONS THROUGH THE INTRODUCTION OF SYNTHETIC DNA. MICROBES ARE BEING GENETICALLY PROGRAMMED TO ACT AS NUTRIENT-PRODUCING BIOFERTILIZERS, PLANT BIOSTIMULANTS THAT ENHANCE FITNESS, PATHOGEN CONTROL AGENTS, AND IN SITU SENSORS FOR CHEMICALS AND SIGNALS. MICROBES NATURALLY EXCHANGE GENETIC INFORMATION, AND SO AS THESE TECHNOLOGIES ARE PUT INTO THE SOIL, SYNTHETIC DNA WILL BE TRANSFERRED FROM ENGINEERED MICROBES TO THE WILD MICROBIAL COMMUNITIES IN THESE SOILS. THIS UNINTENDED TRANSFER OF SYNTHETIC DNA WILL CAUSE DISRUPTION TO THE NATURAL COMMUNITY, WHICH IN TURN CAN ALTER THE PRODUCTIVITY OF THE SOIL, ALTER THE SOIL ECOSYSTEM, OR HAVE LARGER ENVIRONMENTAL EFFECTS. BIOCONTAINMENT STRATEGIES THAT PREVENT RELEASE OF SYNTHETIC DNA ARE BEING INVENTED, BUT THESE ARE NOT PERFECT AND EVEN THE MOST EFFECTIVE TOOLS STILL RELEASE SYNTHETIC DNA. WHILE NEGATIVE IMPACTS OF SYNTHETIC DNA TRANSFER ARE ANTICIPATED, A MAJOR CHALLENGE WITH UNDERSTANDING THE RISKSOF THESE TECHNOLOGIES IS THE LACK OF SIMPLE TOOLS FOR MONITORING GENE TRANSFER FROM SYNTHETIC MICROBES TO WILD MICROBIAL COMMUNITIES.OUR GOAL IS TO DEVELOP A NEW FRAMEWORK FOR MONITORING AND PREDICTING GENE TRANSFER FROM SYNTHETIC MICROBES TO ENVIRONMENTAL MICROBIAL COMMUNITIES IN SOIL. FIRST, WE WILL CREATE A NEW TOOL CALLED CATALYTIC RNA (CAT-RNA) THAT IS ABLE TO RECORD GENE TRANSFER EVENTS WITHIN SOIL. CAT-RNA IS AN RNA-BASED MEMORY DEVICE THAT IS EMBEDDED WITHIN A SYNTHETIC DNA SEQUENCE AND DESIGNED TO RECORD EACH HOST THAT THE SYNTHETIC DNA ENTERS. THESE RECORDINGS ARE THEN READ OUT BY RNA SEQUENCING. CAT-RNA WILL THEN BE USED IN SYNTHETIC SOILS TO MEASURE HOW DIFFERENT SOIL PROPERTIES EFFECT GENE TRANSFER RATES. THIS WILL USE INNOVATIVE SYNTHETIC SOILS THAT ARE TUNABLE FOR THEIR PROPERTIES (E.G. PH, PARTICLE SIZE). WE WILL ALSO USE CAT-RNA IN A WILD TEXAS SOIL AND MEASURE THE EFFECTIVENESS OF A BIOCONTAINMENT STRATEGY. MEASUREMENTS PERFORMED IN SOILS WILL BE USED TO EXTEND PREDICTIVE MODELS OF GENE TRANSFER, WHICH IN THE FUTURE CAN BE USED TO PREDICT RISK OF GENE TRANSFER FOR A GIVEN ENVIRONMENT AND SYNTHETIC DNA.THROUGH COMPLETION OF THIS PROPOSAL, WE WILL CREATE NEW TOOLS THAT ALLOWS RESEARCHERS TO STUDY GENE TRANSFER IN SOILS. BY USING THESE TOOLS, WE WILL RESOLVE DEEPER UNDERSTANDING OF THE RISK OF GENE TRANSFER IN DIFFERENT SOIL TYPES. USING THIS UNDERSTANDING, WE WILL CREATE PREDICTIVE MODELS THAT CAN BE USED BY REGULATORS TO ASSESS THE RISK OF GENE TRANSFER FOR A GIVEN SOIL TYPE. FINALLY, CAT-RNA CAN BE USED TO ASSESS THE EFFECTIVENESS OF BIOCONTAINMENT STRATEGIES. TOGETHER ALL OF THESE WILL INCREASE THE SAFETY OF SYNTHETIC MICROBE TECHNOLOGIES AND PROVIDE DATA TO ALLOW FOR INFORMED POLICY DECISIONS ON THE REGULATION OF THESE TECHNOLOGIES.