Optimization of mRNA Display for Identifying Peptides that Bind with Thymidylate Synthase RNA

In vitro display technologies are powerful tools for isolating polypeptide binders against various target molecules. The mRNA display approach to in vitro protein selection is based upon the puromycin-mediated formation of a covalent bond between an mRNA and its gene product. In this paper, we use mRNA display to isolate peptides that bind thymidylate synthase (TS) RNA with high affinity from a large peptide library (>1013 different sequences). The selection scheme was constructed, and the experimental conditions, including library synthesis, formation and isolation of RNA-peptide fusions were optimized. mRNA display could play a key role in the discovery of peptides with desired functions.