Development of RNA-based translation modulation tools for synthetic biology applications
Hakstol, Rhys J.
University of Lethbridge. Faculty of Arts and Science
Lethbridge, Alta. : Universtiy of Lethbridge, Department of Mathematics and Computer Science
The ability to reliably regulate translation of heterologous proteins is of great interest for diverse applications in the field of biological and metabolic engineering. RNA-based tools to control ribosome-dependent synthesis of proteins within E. coli were developed. First, adaptation of the well-studied RNA-IN and RNA-OUT system based on E. coli Tn10 to reliably repress translation regardless of coding sequence context was performed. Using an invariant sequence within the RNA-IN/OUT interacting region does not interfere with the function of the RNAs involved. The same RNA-IN/RNA-OUT system was adapted to activate the translation of a specific target mRNA. Regulation of translation extends to processing of polyproteins. In eukaryotes this can be accomplished by viral 2A peptides, avoiding reinitiating at a downstream start codon but has not been observed in bacteria. Therefore, a strategy for developing libraries of 2A peptides with potential activity across an array of organisms was developed.
2A-peptides , biochemistry , gene expression regulation , novel trans-acting RNAs , prokaryotic gene expression , synthetic biology , Dissertations, Academic