Genetically designed microbes for bioimaging and biosensing

Abstract

The advantageous approach to the utilization of the microbes for bioimaging and biosensing underlies under their active motility and self-propulsion characteristics besides their easy bioengineering feature to gain multi-functional activities. The emerging developments make use of microorganisms as therapeutic agents in disease diagnosis and treatment. The dynamic nature of the habitat forces the microorganisms to acclimate themselves to changing living conditions via evolving exclusive bio-functionalities for their survival. Therefore, the living microorganisms producing functional materials serve as a biohybrid system with unprecedented potential for enhancing the detection of a disease biomarker molecule or meeting the great need in cancer diagnosis. The synthetic biology approach, a multidisciplinary field of science, gives the ability to engineer and modulate the microorganisms to redesign existing natural pathways, resulting in the gain of the desired function. Inspiring form nature, the biomineralization of iron-oxide materials is demanding for their potential usage in antitumor effect due to their easy modulation, stability, and magnetic properties. Furthermore, the certain respiratory capacities of electrochemically active microbes enable the respiration of diverse inorganic and organic molecules for their survival in redox-stratified environments. The ability of exchanging electrons with electrodes possesses several diverse biotechnological applications like the construction of microbial fuel cells, electro-fermentation, and electro-genetics. In this thesis, the microbes were engineered for their utilization in bioimaging and biosensing applications. Firstly, intracellular and extracellular magnetite accumulating Escherichia coli bacterial cell machineries were constructed as contrast agents for the MRI scanning, promising for a cancer diagnostic. Secondly, the intracellular magnetite accumulating bacterial cells, possessing all the redox reactions that readily take place in their cytoplasm via synthetically produced proteins, were further engineered to improve their targeting capability for breast cancer tumor cells by displaying a certain nanobody on the cell surface. Thirdly, electronic sentinel bacterial cells were designed utilizing the electron transfer modules for extracellular electron consumption by targeted acceptors for their wireless biomonitoring applications upon detecting a disease molecule. The methodologies described in this thesis are envisioned as promising tools for diagnostic applications.