Post-Transcriptionally Modified tRNAs for Enhanced Translation

Abstract

Synthesizing peptides from non-natural amino acids presents challenges such as maintaining a correct reading frame and proper tRNA charging. Flexizyme aids tRNA charging with non-natural amino acids, often using in-vitro transcribed tRNAs. In-vivo transcribed tRNAs, with post-transcriptional modifications, hypothetically offer better stability. Stability for tRNAfMET was assessed via Tm values, indicating higher stability in in-vivo transcribed tRNAs. However, Hibit luminescence assays showed no significant translation efficiency difference between in-vitro and in-vivo transcribed tRNAs. In conclusion, post-transcriptional modifications may not significantly enhance flexizyme reactions. Future steps involve identifying limiting factors in the system or tRNAs.