Translation of 2′-modified mRNAin vitroandin vivo | Zendy

Helle Aurup | Zendy; Angela Siebert | Zendy; Fritz Benseler | Zendy; David M. Williams | Zendy; Fritz Eckstein | Zendy

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Translation of 2′-modified mRNAin vitroandin vivo

Author(s) -

Helle Aurup,

Angela Siebert,

Fritz Benseler,

David M. Williams,

Fritz Eckstein

Publication year - 1994

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/22.23.4963

Subject(s) - reticulocyte , biology , luciferase , messenger rna , microbiology and biotechnology , in vivo , translation (biology) , biochemistry , nucleoside , in vitro , protein biosynthesis , methionine , amino acid , gene , transfection

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.

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