We have previously demonstrated the storage and retrieval of increased information within a semi-synthetic organism (SSO) through use of an unnatural base pair (UBP) within its DNA. The UBP can be replicated, transcribed into corresponding unnatural codons and anticodons of mRNA and tRNA, allowing for decoding at the ribosome to incorporate non-canonical amino acids (ncAAs) into protein. We continue to optimize our SSO by exploring various deoxy- and ribonucleotide triphosphates to further understand how DNA and RNA polymerases recognize and respond to unnatural triphosphates to obtain <i>in vivo</i> structure activity relationships (SARs) analyses of these nucleotides. We found a large variety of unnatural ribonucleotides that can be transcribed into unnatural RNA and the used to mediate synthesis of ncAA-containing proteins. These results identified the most optimal UBP for information storage and unnatural ribonucleoside triphosphates for its retrieval. The optimized SSO can now produce proteins containing high-density ncAAs.

Optimization of replication, transcription, and translation in a semi-synthetic organism

2019 Fall National Meeting

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