Abstract
To study biological processes in various cell types using unnatural amino acids, we developed new methods for efficient incorporation of unnatural amino acids into proteins in yeast, mammalian cells, and stem cells. A general method was developed to express orthogonal prokaryotic tRNAs in eukaryotic cells using special polymerase III promoters. Mutant synthetases specific for unnatural amino acids were evolved in yeast and transferred for use in mammalian cells and stem cells. Other factors that decrease the incorporation efficiency were also addressed. Supported by the Searle Scholar Program and the Beckman Young Investigator Program.
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