pairs with G73 discriminator base of tRNA
Dinitro-flexizyme-C (dFx-C) is a 46-nucleotide catalytic RNA belonging to the flexizyme family of in vitro-selected aminoacyl-tRNA synthetase ribozymes. Flexizymes recognize tRNA exclusively through base-pairing with the 3′-CCA terminus and the discriminator base at position 73; dFx-C is specifically configured to pair with tRNAs bearing an adenosine at the G73 discriminator position. As a dinitro-flexizyme variant, dFx-C accepts amino acid substrates activated as 3,5-dinitrobenzyl esters, expanding the chemical scope of chargeable substrates beyond those accessible to the original flexizyme or enhanced flexizyme (eFx). dFx-C is a core component of the Flexible In Vitro Translation (FIT) system, in which flexizymes charge nonproteinogenic amino acids onto tRNAs whose anticodons are reassigned to vacant codons, enabling ribosomal synthesis of sequence-defined nonstandard peptides including those containing N-methyl amino acids, D-amino acids, and backbone-modified residues.
Applications:
1) Zhang, J., & Ferré-D'Amaré, A.R. (2014). A Flexible, Scalable Method for Preparation of Homogeneous Aminoacylated tRNAs. Methods in Enzymology, 105–113
2) Goto, Y., Katoh, T., & Suga, H. (2011). Flexizymes for genetic code reprogramming. Nature Protocols, 6(6), 779–790
3) Passioura, T., & Suga, H. (2014). Reprogramming the genetic code in vitro. Trends in Biochemical Sciences, 39(9), 400–408
4) Xiao, H., Murakami, H., Suga, H., & Ferré-D'Amaré, A.R. (2008). Structural basis of specific tRNA aminoacylation by a small in vitro selected ribozyme. Nature, 454(7202), 358–361
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