Position Specific Incorporation of Nonnatural Amino Acids into Proteins In Vivo

Abstract

Incorporation of nonnatural amino acids into proteins in cells opens up new vistas for protein study, protein engineering and cell engineering. In this study, to enable incorporation of various nonnatural amino acids into protein in mammalian cells, we attempted to introduce nonnatural aminacyl-tRNAs into the cells. We designed TatEF-Tu, a new carrier for aminoacyl-tRNA delivery into cells. TatEF-Tu is a fusion protein of TAT cell-penetrating peptide and elongation factor Tu (EF-Tu). TatEF-Tu can bind aminoacyl-tRNAs carrying nonnatural amino acids, protect them from hydrolysis, and deliver them into cells.