ECSUB · Encoded Cellular Synthesis of Unnatural Biopolymers

We are building a parallel and independent (orthogonal) translational machinery for the encoded biosynthesis of unnatural polymers in living cells. The orthogonal translation system has many potential applications beyond those possible with the natural translation system: I propose to use it: 1) To expand the chemical scope of monomers that can be polymerized by the ribosome in living cells, allowing the incorporation of monomers with unnatural backbones into proteins; 2) To increase the efficiency of in vivo unnatural amino acid mutagenesis via amber suppression, so that no truncated protein is produced and multi-site incorporation of unnatural amino acids is possible; 3) To create probes of protein function for use in vivo; 4) To free numerous codons for simultaneous encoding of multiple distinct unnatural monomers, and to experimentally explore alternate genetic codes; 5) To explore the evolution of encoded unnatural polymers toward new cellular functions.