TetR-binding aptamer as a versatile regulatory element

Adam Artur Mol
Synthetic biology explores the means of redesign and fabrication of existing biological systems or the de novo design and generation of biological components that are entirely new to nature. The main focus of the relatively new and dynamic discipline is to develop programmable genetic regulatory systems. Precise, reversible and temporary control of gene expression as well as in-depth understanding of fundamental genetics are crucial for the programming of new genetic circuits. RNA represents
more » ... of the most powerful substrates in the engineering of biological systems, as it is versatile, designable and easily characterizable. Among the diverse functions of RNA molecules, their role as natural riboswitches has predominantly inspired researchers to design synthetic RNA-based regulators. Most of RNA devices contain a sensor element, an aptamer domain, which recognizes small molecules or protein ligands with high specificity and affinity, and an expression platform, controlling gene expression via various mechanisms. Generally, binding of a specific ligand to the aptamer domain stabilizes the RNA molecule or causes conformational changes in its structure; these further regulate transcription, translation and mRNA processing and degradation. Engineered RNA-based devices have already demonstrated multiple applications in synthetic biology. However, their implementation was mostly validated in bacteria and yeast, while mammalian synthetic biology has lagged behind. Splicing of pre-mRNAs is an essential process in human cells that generates a diverse proteome through networks of coordinated splicing events and offers an additional layer of control for synthetic RNA devices. The reprogrammed removal of intronic sequences could provide a novel approach for the development of gene therapies to tackle disease phenotypes. For this purpose, it is necessary to design tools that allow precise and timely control of the splicing mechanisms. In the first research project described in the presented doctoral thesis, a versatile and h [...]
doi:10.25534/tuprints-00011845 fatcat:k65wck6ypnab3mhxf7ivdxpy6e