Next generation tools for studying UPEC pathogenesis
Speaker: MD/PhD Swaine Chen, Singapore NRF Fellow, National University of Singapore and Genome Institute of Singapore, Singapore.
Title: "Next generation tools for studying UPEC pathogenesis"
Room: Major Groove
Hosts: Fredrik Almqvist and Sven Bergström
Abstract: Urinary tract infections (UTIs) are extremely common infections affecting half of all women and contributing greatly to antibiotic prescriptions and therefore bacterial antibiotic resistance. Most UTIs are caused by uropathogenic Escherichia coli (UPEC). The study of how UPEC cause UTIs is greatly facilitated by many genetic and molecular tools available for E. coli in general. However, these tools are usually developed in and for cloning or lab-adapted strains of E. coli, and they are sometimes not usable or not efficient in disease-causing clinical isolates such as UPEC.
My lab has developed two new tools with clinical strains in mind to improve our ability to study UTI. The first improves our ability to manipulate the UPEC chromosome. We have developed a general and modular negative selection (counterselection) system that functions without optimization in multiple clinical isolates of E. coli and Salmonella. Furthermore, this system functions up to 1000x better than all other reported systems in lab strains of E. coli. This system now enables the convenient creation of definitive genetic constructs directly in UPEC.
The second project improves our ability to detect UPEC during infection. We have designed a new GFP protein which we term vGFP that demonstrates a 30-50% improvement in bulk brightness while simultaneously enabling rational control of dimerization state. vGFP is therefore a better chromosomal reporter than other GFP variants and should improve our detection sensitivity of UPEC, which is paramount for tracking UPEC as they move through minor niches during infection.
Together these tools improve our ability to translate our mouse studies into human disease. I will conclude with our plans for performing niche-specific, single-cell resolved, simultaneous host and pathogen transcriptional profiling in direct UTI samples. In the future, this will hopefully allow us to do a full genomic translation of our lab studies into knowledge about human disease.