To defeat tuberculosis, we need new biotechnology. Our work adds 4 new tools to the anti-TB medical armamentarium. Detect - a CRISPR-based biosensor delivered by phage and sequence-specific for antibiotic resistance. Target - an E. coli model hosting an essential mycobacterial metabolic pathway that could simplify drug screening. Infiltrate - an E. coli strain capable of entering infected macrophages and lysing mycobacteria. Sabotage - a non-lytic phage that spreads horizontally in a bacterial population and expresses an siRNA to knock down antibiotic resistance.
Diagnosing antibiotic resistance can improve and accelerate treatment. We propose a phage-delivered, CRISPR-based system that cuts specific DNA sequences and detects the presence of resistance genes due to the resulting DNA damage that is reported with a color output.
M. tuberculosis grows slowly and is hard to study in the lab. We have transferred an essential mycobacterial metabolic pathway to E. coli, where it is easy to screen for targeted small-molecule inhibitors.
An effective TB therapy must reach mycobacteria inside lung macrophages. In this system, E. coli express listeriolysin O (LLO) to enter the macrophage cytosol and Trehalose Dimycolate Hydrolase (TDMH) to degrade the pathogen's membrane.
Totally drug-resistant TB (TDR-TB) is an established and growing problem. We have created a phage vector that delivers an siRNA capable of sabotaging drug resistance and restoring sensitivity. By reducing the fitness burden of our construct, we can promote its spread in a population.