Flexizyme as a versatile tRNA acylation catalyst and the application for translation - RNA

Flexizyme as a versatile tRNA acylation catalyst and the application for translation
2006
Hiroshi Murakami1, Atushi Ohta2, Yuki Goto1, Yusuke Sako2 and Hiroaki Suga1,2
1 Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan, 2 Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyou-ku, Tokyo 113-8656, Japan
Nucleic Acids Symposium Series 2006
Here we describe a de novo tRNA acylation system consisting of artificial ribozymes. This system, the flexizyme (Fx) system, allows for the preparation of acyl-tRNAs with almost no limitation on the use of a variety of amino/hydroxy acids and tRNAs. To demonstrate its utility, we have carried out protein synthesis involving site-specific incorporation of nonnatural amino and hydroxy acids via amber or programmed frame-shift suppressions. We have also demonstrated peptide synthesis involving multiple nonnatural amino acids via sense codon suppression. The combination of the Fx system and appropriate cell-free translation is a powerful and flexible tool for mRNA-encoded synthesis of nonnatural polypeptides.
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