A Foundational Synthetic Biology Toolbox for Engineering Human Gut Microbiota towards Enhancing Warfighter Capabilities
Abstract
PROJECT SUMMARY The overarching goal of the proposed research is to develop foundational synthetic biology technologies that enable the engineering of gut bacteria and probiotics to modulate gutbrain- microbiota interactions in naval personnel for improving stress resilience and operational readiness and performance. Microbes of the gastrointestinal tract interact with the host enteric nervous system and immune system to modulate host response to various stressors both environmental (e.g. temperature, day-night cycles) and biological (e.g. pathogens), as well as participate in essential metabolism of foods and medicine. Deployment of genetic sensors, actuators, and engineered circuitry to modify these gut bacteria in various synthetic biology applications require state-of-the-art capabilities in genome engineering, plasmid maintenance, and DNA transformation. Currently, large technological gaps need to be filled to enable efficient and facile engineering of human-associated microbial communities. Here, we propose to address these key challenges. Specifically, we propose to develop a new genome engineering method using CRISPR-transposons, a new plasmid capture technology for developing novel and better plasmid vectors, and a new class of DNA transformation methods for gut bacteria using claybased nanoporation. The research activities are broken down into three major thrusts. First, we aim to develop a new genome engineering approaching using a CRISPR-Cas9-transposase fusion to site-specifically insert programmable sequences into given a genome for targeted editing in gut microbes, first in aerobic E. coli as a model but then in Bacteroides species. Second, we aim to develop methods to capture and reengineer mobile plasmid vectors from natural gut microbes to expand the essential repertoire of plasmid backbones that for the foundation for genetic and genomic engineering. Third, we aim to develop a new transformation method using “nanopiercing” with naturally occurring sepiolite clay to introduce genetic material into gut microbiota. The proposed research has the potential to become a transformative platform for next-generation human microbiome augmentation and engineering for various naval-relevant applications.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Aug 12, 2016
- Source ID
- N000141512704
Entities
People
- Harris H. Wang
Organizations
- Office of Naval Research
- Trustees of Columbia University in the City of New York
- United States Navy