Preparation of materials for flexizyme reactions and genetic code reprogramming
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
Posted 12 May, 2011
Preparation of materials for flexizyme reactions and genetic code reprogramming
Posted 12 May, 2011
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
© Research Square 2021