A facile system for encoding unnatural amino acids in mammalian cells.
A shuttle system has been developed to genetically encode unnatural amino acids in mammalian cells using aminoacyl-tRNA synthetases (aaRSs) evolved in E. coli. A pyrrolysyl-tRNA synthetase (PylRS) mutant was evolved in E. coli that selectively aminoacylates a cognate nonsense suppressor tRNA with a photocaged lysine derivative. Transfer of this orthogonal tRNA-aaRS pair into mammalian cells made possible the selective incorporation of this unnatural amino acid into proteins.
Chen
Peng R
PR
Department of Chemistry, The Scripps Research Institute, 10550 Torry Pines Road, La Jolla, CA 92037, USA.
Groff
Dan
D
Guo
Jiantao
J
Ou
Weijia
W
Cellitti
Susan
S
Geierstanger
Bernhard H
BH
Schultz
Peter G
PG
D000596:Amino Acids / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001105:Archaea / Q000235:genetics; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005815:Genetic Code; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry* / Q000235:genetics
D000596:Amino Acids; D011506:Proteins; D000604:Amino Acyl-tRNA Synthetases; C456839:pyrrolysine; D008239:Lysine