Developing smRNA FISH for the identification of antibiotics-tolerant persister bacteria
Evelina Tutucci (evelina.tutucci[at]vu.nl)
Wilber Bitter (firstname.lastname@example.org)
Frank Bruggeman (email@example.com)
Multi-drug-resistant, pathogenic bacteria threaten human health. They have acquired DNA mutations that make them insensitive to antibiotics. Pathogenic bacteria are also successful in dealing with antibiotics, and the human immune system, because they often can switch to a non-growing, antibiotics-tolerance state, called the ‘persister’ state. Persister cells are formed from growing bacteria, can switch back to the growing state and coexist with growing cells in cell cultures. Since persister cells are not growing, they can withstand the effects of antibiotics that kill growing cells. It has been shown that growing cells can switch into persister cells, via so-called toxin-antitoxin systems. Such systems have been discovered in Escherichia coli and have since been found in many pathogenic bacteria, including Pseudomonas aerogenosa and Mycobacterium tuberculosis. Understanding how persister cells form is an important research topic.
Aim of this project
Since persister cells derive from growing cells and coexist with them in growing cell culture, we require single-cell methods to identify persister cells. One promising method is to determine the relative number of toxin and antitoxin transcripts in single cells, as it is believed that an excess of toxins over antitoxins turn growing cells into persister cells. The aim of this project is develop single-molecule RNA FISH, which allows for counting of the number of transcripts in single cells, for identification of persister cells. First in Escherichia coli and subsequently in Mycobacterium segmatus, a relative of M. tuberculosis.
Methods and techniques
fluorescence microscopy, molecular biology, bacterial cell cultivation, RNA methods, smRNA FISH.
During this project you become familiar with state-of-the-art methods of microbiology, molecular biology and fluorescence microscopy that are used throughout molecular biology labs in hospitals, academia and industry. Since smRNA FISH is not routinely used in such labs, the expertise which you acquire during this project will count on the job market, regardless whether this concerns a fundamental or applied job.
Number of positions available
Duration of project
From July 2024