Antibiotic resistance is an urgent global threat resulting in millions of infections and thousands of deaths in the United States alone. This involves bacteria surviving, proliferating, and spreading their resistant mechanisms, despite antibiotic efforts. This project examines a novel target pathway: trans-translation, an essential ribosome rescue process in bacterial pathogens. A Caulobacter model is utilized for cloning and ribosome profiling methodologies to gain crucial insight into each step of trans-translation for optimizing novel antibiotic development.