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Method Article
Preparation of materials for flexizyme reactions and genetic code reprogramming
Hiroaki Suga
Department of Chemistry, Graduate School of Science, The University of Tokyo
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Genetic code reprogramming is a method for the reassignment of arbitrary codons from proteinogenic amino acids to non-proteinogenic ones, and thus specific sequences of non-standard peptides can be ribosomally expressed according to their mRNA templates. We have developed a protocol that facilitates the genetic code reprogramming using flexizymes integrated with a custom-made in-vitro translation apparatus, referred to as the flexible in-vitro translation (FIT) system. Flexizymes are flexible tRNA acylation ribozymes that enable the preparation of a diverse array of non-proteinogenic acyl-tRNAs. These acyl-tRNAs read vacant codons created in the FIT system, yielding the desired non-standard peptides with diverse exotic structures, such as N-methyl amino acids, D-amino acids and physiologically stable macrocyclic scaffolds. We here describe a protocol for the preparation of DNA templates for flexizymes, tRNAs, microhelix RNA, and mRNAs that are used for genetic code reprogramming with flexizymes.
Keywords
Translation, Ribosome, Genetic code, tRNA, Aminoacylation, Non-proteinogenic amino acids, Ribozyme
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This is a preprint. It has not completed peer review.
Method Article
Preparation of materials for flexizyme reactions and genetic code reprogramming
Hiroaki Suga
Department of Chemistry, Graduate School of Science, The University of Tokyo
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 12 May, 2011
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Subject Areas
Biochemistry
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Genetic code reprogramming is a method for the reassignment of arbitrary codons from proteinogenic amino acids to non-proteinogenic ones, and thus specific sequences of non-standard peptides can be ribosomally expressed according to their mRNA templates. We have developed a protocol that facilitates the genetic code reprogramming using flexizymes integrated with a custom-made in-vitro translation apparatus, referred to as the flexible in-vitro translation (FIT) system. Flexizymes are flexible tRNA acylation ribozymes that enable the preparation of a diverse array of non-proteinogenic acyl-tRNAs. These acyl-tRNAs read vacant codons created in the FIT system, yielding the desired non-standard peptides with diverse exotic structures, such as N-methyl amino acids, D-amino acids and physiologically stable macrocyclic scaffolds. We here describe a protocol for the preparation of DNA templates for flexizymes, tRNAs, microhelix RNA, and mRNAs that are used for genetic code reprogramming with flexizymes.
Figure 1
Figure 2
Figure 3