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bioRxiv · 10.1101/2024.12.12.627959

Engineering a ribozyme with aminoacyl-tRNA synthetase activity

Gan, G.·Chen, J.·Manuel, G.·Corn, R. M.·Luptak, A.·Golden, B.
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Abstract

A ribozyme that can charge a tRNA with amino acids and discriminate between cognate and non-cognate tRNAs is of interest because an RNA with this ability may have been a critical for translation in the transition from the RNA world. In addition, it could provide a tool for incorporating non-canonical amino acids for biotechnology applications. Here, we rationally engineer a ribozyme by fusing a tRNA binding module derived from a T-box riboswitch with a catalytic module (a flexizyme) to generate a ribozyme that can amino acylate a target tRNA. We demonstrate that this ribozyme be readily redesigned to alter tRNA specificity. This ribozyme is compatible with an in vitro translation system and could be used to recode a protein sequence to site-specifically incorporate a non-canonical amino acid.

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Gan, G., Chen, J., Manuel, G., Corn, R. M., Luptak, A., Golden, B.. 2024-12-12. Engineering a ribozyme with aminoacyl-tRNA synthetase activity. https://doi.org/10.1101/2024.12.12.627959

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