Grantee Research Project Results
2024 Progress Report: Developing Microbial Biocontainment Strategies and Their Assessment Methods
EPA Grant Number: R840205Title: Developing Microbial Biocontainment Strategies and Their Assessment Methods
Investigators: Moon, Tae Seok , Parker, Kimberly M
Institution: Washington University in St. Louis
EPA Project Officer: Callan, Richard
Project Period: July 1, 2021 through May 10, 2025
Project Period Covered by this Report: July 1, 2023 through June 30,2024
Project Amount: $744,262
RFA: Assessment Tools for Biotechnology Products (2020) RFA Text | Recipients Lists
Research Category: Safer Chemicals , Chemical Safety for Sustainability
Objective:
Our long-term vision is to enable the successful development and assessment of biocontainment strategies to understand and prevent the escape of genetically engineered microbes (GEMs) in the environment. To this end, we aim to (1) develop a novel genetic “suicide” circuit that turns on only after the designed “mission” of a GEM is accomplished and (2) assess its function and stability under environmentally-relevant conditions.
Progress Summary:
In this last period, we explored the effect of additional environmental factors like temperature on the effectiveness of kill switch, measured using GEM escape rate. We also developed approaches to measure the persistence of genetic markers of biocontained GEMs in environmental samples. In addition, we developed biosensors that can be used for potential biocontainment circuits and demonstrated kill switch functionality that is triggered only after bacteria consume toxic compounds such as phenol in the environment. Importantly, we developed the antibiotic resistance gene-free plasmid (ARGFP)-based cloning method to potentially reduce antibiotic resistance spread caused by horizontal gene transfer of antibiotic resistance genes that have been disposed of and originated from research laboratories all over the world for many decades. This ARGFP-based method, if broadly adopted, will revolutionize the current laboratory practice of biowaste treatment.
Future Activities:
During the next period, we will continue our efforts to test kill switches under environmental systems through completing remaining steps to revise our manuscript demonstrating elevated escape rates in environmental matrices and by finalizing experiments exploring the persistence of genetic markers from biocontained GEMs. In addition, Parker and her team plan to prepare a perspective piece introducing GEMs, their applications, and their risk assessment to environmental engineers, as well as to participate in synthetic biology focused sessions at upcoming conferences. We will also continue to develop and assess aromaticdependent kill switches in soil bacteria such as Rhodococcus and Pseudomonas strains. Specifically, we will prepare or revise the papers based on reviewers’ comments. Notably, it will include the iterative design-build-test-learn cycle to ensure the genetic stability of kill switches under real environmental conditions.
Journal Articles on this Report : 1 Displayed | Download in RIS Format
| Other project views: | All 9 publications | 8 publications in selected types | All 8 journal articles |
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Xi C, Ma Y, Amrofell MB, Moon TS. Manipulating the molecular specificity of transcriptional biosensors for tryptophan metabolites and analogs. Cell Reports Physical Science 2024;5(10). |
R840205 (2024) |
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Supplemental Keywords:
genetically modified organism; biosafety; cleanup; genetic circuit.Relevant Websites:
Progress and Final Reports:
Original AbstractThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.