Synthesis and use of anti-reverse mRNA cap analogues

Details Synthesis and use of anti-reverse mRNA cap analogues Synthesis and use of anti-reverse mRNA cap analogues

2006-07-11

2006-07-11

Lilling, Herbert J. (Department: 1651)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

C435S005000, C435S006120, C435S091500, C435S091100, C435S091510, C536S023100, C536S023200, C536S025400

Reexamination Certificate

active

07074596

ABSTRACT:
The ability to synthesize capped RNA transcripts in vitro has been of considerable value in a variety of applications. However, one-third to one-half of the caps have, until now, been incorporated in the reverse orientation. Such reverse caps impair the translation of in vitro-synthesized mRNAs. Novel cap analogues, such as P1-3′-deoxy-7-methylguanosine-5′P3-guanosine-5′triphosphate and P1-3′-O,7-dimethylguanosine-5′P3-guanosine-5′triphosphate, have been designed that are incapable of being incorporated into RNA in the reverse orientation. Transcripts produced with SP6 polymerase using “anti-reverse” cap analogues were of the predicted length. Analysis of the transcripts indicated that reverse caps were not formed. The in vitro translational efficiency of transcripts with the novel “anti-reverse” cap analogues was significantly higher than that of transcripts formed with conventional caps.

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