Automated orthogonal tRNA generation.

ABSTRACT: The ability to generate orthogonal, active tRNAs-central to genetic code expansion and reprogramming-is still fundamentally limited. In this study, we developed Chi-T, a method for the de novo generation of orthogonal tRNAs. Chi-T segments millions of isoacceptor tRNA sequences into parts and then assembles chimeric tRNAs from these parts. Chi-T fixes the parts, containing identity elements, and combinatorially varies all other parts to generate chimeric sequences. Chi-T also filters the variable parts and chimeric sequences to minimize host identity elements. We show here that experimentally characterized orthogonal tRNAs are more likely to have predicted minimum free energy cloverleaf structures, and Chi-T filters for sequences with a predicted cloverleaf structure. We report RS-ID for t

SUBMITTER: Spinck M 

PROVIDER: S-EPMC12037408 | biostudies-literature | 2025 May

REPOSITORIES: biostudies-literature