Tools4Cells: Harnessing the plasmidome for environmental engineering of microbial communities using synthetic biology

About This Grant

Microbes share information through exchanging DNA elements called plasmids, which are critical for how microbes adapt and evolve new abilities. Researchers use plasmids as tools to give microbes new instructions, like producing medicines or cleaning up pollutants. However, just as software only works on certain types of hardware, different plasmid types can only function in specific types of microbes. While a vast variety of plasmids exist in nature, researchers primarily use a select few, which limits our ability to program different microbes for societal benefits. To overcome this challenge, this project will develop new methods to isolate, study, and use different plasmids. This research will be used to expand the current plasmid toolbox from a few plasmids to hundreds and potentially even thousands, which will enhance our ability to program microbes for beneficial purposes. The beneficial impact of this research will be to expand our ability to address grand challenges using microbes related to public health, food and water security, and energy transitions. In addition, broader outcomes will include the development of new training resources, facilitation of workshops to disseminate research innovations, and the creation of new summer research opportunities for high-school students and community college students. The goal of this research is to expand the available plasmids for researchers to use as tools by establishing a high-throughput platform for isolating and engineering plasmids and characterizing their properties (i.e., host range, stability, connectivity). Novel methods will be developed to achieve this research. First, more efficient and effective methods for isolating DNA plasmids from environmental microbes will be established. Second, an innovative synthetic biology tool will be established to measure the key properties, such as host range, of these novel DNA plasmids. The new plasmid tools will then be used as resources to study the genetic determinants of plasmid host range and plasmid stability in microbial communities. Additionally, we will demonstrate the use of these plasmids as tools to introduce a non-native catabolic gene for phenol pollutant biodegradation into a microbial community, and assess its impact on pollutant biodegradation rates. This project is supported by the Systems and Synthetic Biology Cluster of the Division of Molecular and Cellular Biosciences. 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.