title: Direct and specific chemical control of eukaryotic translation with a synthetic RNA-protein interaction.
abstract: Sequence-specific RNA-protein interactions, though commonly used in biological systems to regulate translation, are challenging to selectively modulate. Here, we demonstrate the use of a chemically-inducible RNA-protein interaction to regulate eukaryotic translation. By genetically encoding Tet Repressor protein (TetR)-binding RNA elements into the 5'-untranslated region (5'-UTR) of an mRNA, translation of a downstream coding sequence is directly controlled by TetR and tetracycline analogs. In endogenous and synthetic 5'-UTR contexts, this system efficiently regulates the expression of multiple target genes, and is sufficiently stringent to distinguish functional from non-functional RNA-TetR interactions. Using a reverse TetR variant, we illustrate the potential for expanding the regulatory properties of the system through protein engineering strategies.
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Goldfless
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Belmont
authors.forename: Alexandra M
authors.initials: AM
authors.lastname: de Paz
authors.forename: Jessica F
authors.initials: JF
authors.lastname: Liu
authors.forename: Jacquin C
authors.initials: JC
authors.lastname: Niles
mesh_terms: D020121:5' Untranslated Regions; D000818:Animals; D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000378:metabolism; D002474:Cell-Free System; D005786:Gene Expression Regulation; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis; D011817:Rabbits; D012097:Repressor Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics
chemical_list: D020121:5' Untranslated Regions; D052157:Aptamers, Nucleotide; D012097:Repressor Proteins; C034811:tetracycline resistance-encoding transposon repressor protein