Preparation of translationally competent tRNA by direct chemical acylation.
Nonsense codon suppression for unnatural amino acid incorporation requires the preparation of a suppressor aminoacyl-tRNA. Chemical acylation strategies are general but inefficient and arduous. A recent report (J. Am. Chem. Soc. 2007, 129, 15848) showed acylation of RNA mediated by lanthanum(III) using amino acid phosphate esters. The successful implementation of this methodology to full-length suppressor tRNA is described, and it is shown that the derived aminoacyl-tRNA is translationally competent in Xenopus oocytes.
Duffy
Noah H
NH
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
Dougherty
Dennis A
DA
D000215:Acylation; D000818:Animals; D018389:Codon, Nonsense / Q000235:genetics; D007811:Lanthanum / Q000737:chemistry*; D051379:Mice; D015394:Molecular Structure; D009865:Oocytes / Q000378:metabolism*; D012343:RNA, Transfer / Q000138:chemical synthesis* / Q000737:chemistry; D012346:RNA, Transfer, Amino Acyl / Q000138:chemical synthesis* / Q000737:chemistry; D014981:Xenopus / Q000235:genetics / Q000378:metabolism*
D018389:Codon, Nonsense; D012346:RNA, Transfer, Amino Acyl; D007811:Lanthanum; D012343:RNA, Transfer