Identification and development of cyclic peptide libraries containing non-natural amino acids

The aim of this project is to develop a high-throughput screening platform, that uses droplet microfluidics to co-encapsulate and screen a SICLOIPPS (split-intein circular ligation of peptides and proteins) library of 3.2 million cyclic peptides containing multiple non-natural amino acids. Once established, this platform will be used to identify inhibitors of a therapeutically relevant protein-protein interaction. This work builds on previous work form the Tavasoli lab whereby a library of 3.2 million cyclic peptides have been produced in fL double-emulsion droplets. A combination of chemical and biochemical methods will be used to charge tRNA with non-natural amino acids. The product of these reactions will be used in custom in vitro transcription and translation systems, enabling the production of SICLOPPS libraries containing multiple non-natural amino acids.