Development of an improved transposon mutagenesis system for comprehensive genetic analysis of Borrelia burgdorferi

PROJECT SUMMARY/ABSTRACT
Lyme disease, caused by Borrelia burgdorferi, remains a significant public health concern with
limited treatment options. Despite decades of research, our knowledge of the genes it needs for
transmission and infection is largely incomplete, hindering the development of effective therapies,
vaccines, and diagnostic tests. In this proposal we aim to develop innovative high-throughput
genetic tools that will facilitate the study of B. burgdorferi pathogenesis and accelerate Lyme
disease research. We will create a high-efficiency transposon insertional mutagenesis system for
B. burgdorferi, overcoming current limitations in generating comprehensive mutant libraries. This
system will employ a custom-designed transposon delivery plasmid carrying a multifunctional
Himar1 transposon. We will use transposon sequencing (Tn-seq) to conduct genome-wide fitness
screens under various growth conditions, including different carbon sources relevant to the
pathogen's life cycle. This will allow us to identify essential and conditionally essential genes for
B. burgdorferi survival and growth. In addition, we use the transposon insertion library tool and
Tn-seq to perform a comprehensive genetic interaction mapping of an important transcriptional
regulator to begin to define its downstream regulated genes. By creating these genetic tools and
resources, this project will advance the field of Lyme disease research. The methods developed
here will enable rapid, systematic studies of B. burgdorferi gene function and regulation. The
development of these tools will provide insights into B. burgdorferi's genetic regulatory networks
and essential pathways to aid future research on targeted drug development for Lyme disease
prevention and treatment.

Grant Number: 1R03AI192375-01
NIH Institute/Center: NIH
Principal Investigator: Andrew Camilli