Preparation of eukaryotic mRNA having differently methylated adenosine at the 5'-terminus and the effect of the methyl group in translation.
Cap analogues having differently methylated adenosine at 2' and N6 position, m(7)G(5')pppApG which is existed in plant mRNA (plant type), m(7)G(5')pppAmpG (animal type), m(7)G(5')pppm(6)AmpG (mammalian type) and m(7)G(5')pppm(6)ApG (unnnatural type), were synthesized. In order to clarify the function of these methyl groups, luciferase mRNAs having differently methylated adenosine at the 5'-terminus, were successfully prepared by in vitro transcription using the synthesized cap anologues. As the preliminary results of in vitro translation with rabbit reticulocyte lysate and luciferase assay, luciferase mRNA having the mammalian type of cap structure, m(7)G(5')pppm(6)AmpG, was most efficiently translated. In the case of m(7)G(5')pppApG (plant type) efficiency of translation was lowest.
Ishikawa
Masahide
M
Department of Applied Chemistry, Graduate School of Engineering, Saitama Institute of Technology, Fukaya, Saitama 369-0293, Japan. ishikawa@sit.ac.jp
Murai
Ryuta
R
Hagiwara
Hiroyuki
H
Hoshino
Tetsuya
T
Suyama
Kamui
K
D000241:Adenosine / Q000737:chemistry*; D000818:Animals; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D008745:Methylation; D014176:Protein Biosynthesis; D012314:RNA Cap Analogs / Q000138:chemical synthesis / Q000737:chemistry*; D011817:Rabbits
D012314:RNA Cap Analogs; D008156:Luciferases; D000241:Adenosine