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Rational Design of Aptamer-Tagged tRNAs

Department of Life Science, College of Science, Rikkyo University, 3-34-1 Nishi-Ikebukuro, Toshima-ku, Tokyo 171-8501, Japan
Int. J. Mol. Sci. 2020, 21(20), 7793;
Received: 27 August 2020 / Revised: 14 October 2020 / Accepted: 20 October 2020 / Published: 21 October 2020
(This article belongs to the Special Issue Expanding and Reprogramming the Genetic Code 2.0)
Reprogramming of the genetic code system is limited by the difficulty in creating new tRNA structures. Here, I developed translationally active tRNA variants tagged with a small hairpin RNA aptamer, using Escherichia coli reporter assay systems. As the tRNA chassis for engineering, I employed amber suppressor variants of allo-tRNAs having the 9/3 composition of the 12-base pair amino-acid acceptor branch as well as a long variable arm (V-arm). Although their V-arm is a strong binding site for seryl-tRNA synthetase (SerRS), insertion of a bulge nucleotide in the V-arm stem region prevented allo-tRNA molecules from being charged by SerRS with serine. The SerRS-rejecting allo-tRNA chassis were engineered to have another amino-acid identity of either alanine, tyrosine, or histidine. The tip of the V-arms was replaced with diverse hairpin RNA aptamers, which were recognized by their cognate proteins expressed in E. coli. A high-affinity interaction led to the sequestration of allo-tRNA molecules, while a moderate-affinity aptamer moiety recruited histidyl-tRNA synthetase variants fused with the cognate protein domain. The new design principle for tRNA-aptamer fusions will enhance radical and dynamic manipulation of the genetic code. View Full-Text
Keywords: genetic code; anticodon; pseudo-anticodon; amber suppression genetic code; anticodon; pseudo-anticodon; amber suppression
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MDPI and ACS Style

Mukai, T. Rational Design of Aptamer-Tagged tRNAs. Int. J. Mol. Sci. 2020, 21, 7793.

AMA Style

Mukai T. Rational Design of Aptamer-Tagged tRNAs. International Journal of Molecular Sciences. 2020; 21(20):7793.

Chicago/Turabian Style

Mukai, Takahito. 2020. "Rational Design of Aptamer-Tagged tRNAs" Int. J. Mol. Sci. 21, no. 20: 7793.

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