Translation of 2'-modified mRNA in vitro and in vivo.
2'-Fluoro- and 2'-amino-2'-deoxynucleoside triphosphates have been used for in vitro transcription of 2'-modified luciferase mRNA. The 2'-modified deoxynucleoside-containing transcripts were tested for the expression of luciferase in X.Laevis oocytes as well as in rabbit reticulocyte lysate. Only 2'-fluoro-2'-deoxy-adenosine-modified mRNA gave rise to luciferase as shown by SDS gel as well as by enzyme activity measurements in vivo as well as in vitro. 2'-Fluoro-2'-deoxy-pyrimidine nucleoside-modified mRNA did not give rise to luciferase activity. However, they directed incorporation of 35S-labeled methionine into peptide fragments in rabbit reticulocyte lysate indicating premature termination of translation. No or only extremely little of such incorporation could be detected with 2'-amino modified transcripts.
Aurup
H
H
Max-Planck-Institut für experimentelle Medizin, Göttingen, Germany.
Siebert
A
A
Benseler
F
F
Williams
D
D
Eckstein
F
F
D000818:Animals; D001483:Base Sequence; D002474:Cell-Free System; D003854:Deoxyribonucleotides / Q000138:chemical synthesis*; D005786:Gene Expression Regulation; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008715:Methionine / Q000378:metabolism; D008969:Molecular Sequence Data; D009865:Oocytes; D014176:Protein Biosynthesis / Q000502:physiology*; D012315:RNA Caps / Q000096:biosynthesis; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics*; D011817:Rabbits; D014158:Transcription, Genetic; D014982:Xenopus laevis
D003854:Deoxyribonucleotides; D012315:RNA Caps; D012333:RNA, Messenger; D008715:Methionine; D008156:Luciferases