Tabula Rasa: Decoupling genetic information from medium

Abstract

tRNAs are indispensable participants in the translation process. On each tRNA, there is a three-base-pair anticodon that binds with the codon on mRNA, such that only one corresponding amino acid can be added to the peptide chain each time. In synthetic biology, control over the final product generally occurs at the nucleic acid coding level while the translation process is held constant. In this project, we utilized tRNA biology to create a system where translation patterns can be changed in a controllable, inducible manner, decoupling the product from the genetic code. To achieve this goal, we are building a gene circuit composed of a constitutively expressed nonsense mutated GFP, one induced tyrosine amber suppressor tRNA, and one induced histidine amber suppressor tRNA. Currently, we realized a transformation from a mutated GFP Y66X to BFP (GFP Y66H) dependent on a coexpressed histidine suppressor tRNA. This proof of concept demonstrates that plasmid-delivered tRNAs are processed, involved in the translation, and can rescue the fluorescent protein.