The construction of artificial gene circuits for synthetic biology applications necessitates the design of modular and inter-connectable molecular switches. Translational repression switches involve the repression of translation from a target messenger RNA (mRNA) by binding of a ligand to sites located in the 5'-untranslated region (UTR) of that mRNA. Although, previous studies have demonstrated the potential of translational repression switches for the construction of gene circuits, specific guidelines for the construction of novel switches are lacking. Hence, we developed a design strategy to guide the construction of future translational repression switches. Specifically, we designed a genetically encoded switch incorporating two repeats of bovine immunodeficiency virus (BIV) TAR based RNA hairpins in the 5'- UTR that showed 7-fold translational repression with BIV Tat ligand. We characterized the influence of RNA hairpin location, number of hairpin repeats, and the thermodynamic stability of the hairpin on the extent of translational repression. The strategy described in this work could be readily used to further expand the repertoire of these key molecular switches.

Design of a translational repression switch to control gene-expression using BIV Tat peptide-TAR RNA interaction in mammalian cells

247th National Meeting (2014)

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