OLIGONUCLEOTIDE-MEDIATED SENSE CODON REASSIGNMENT

摘要

Sense codon reassignment to unnatural amino acids (uAAs) represents a powerful approach for introducing novel properties into polypeptides. The main obstacle to this approach is competition between the native isoacceptor tRNA(s) and orthogonal tRNA(s) for the reassigned codon. While several chromatographic and enzymatic procedures for selective deactivation of tRNA isoacceptors in cell-free translation systems exist, they are complex and not scalable. The present invention provides oligonucleotides that hybridise to a tRNA of interest when said tRNA is in a folded state, thereby disrupting the function of the tRNA. The present invention also provides the use of these oligonucleotides in methods for sense codon reassignment and methods for incorporation of uAAs into proteins. The approach described herein represents a new direction in genetic code reassignment with numerous practical applications.