Flexizymes as a tRNA acylation tool facilitating 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 nonstandard peptides can be ribosomally expressed according to their mRNA templates. We here describe 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 nonstandard peptides with diverse exotic structures, such as N-acyl groups, D: -amino acids, N-methyl amino acids, and physiologically stable macrocyclic scaffolds. Facility of the protocol allows for a wide variety of applications in the synthesis of new classes of nonstandard peptides with biological functions.
Department of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo, Japan.
Yuki
Y
Goto
Hiroaki
H
Suga
D001483:Base Sequence; D005815:Genetic Code / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D016337:RNA, Catalytic / Q000378:metabolism*; D012343:RNA, Transfer / Q000235:genetics*; D046249:Transfer RNA Aminoacylation
D016337:RNA, Catalytic; D012343:RNA, Transfer