Louisiana State University and Agricultural & Mechanical College

Janusz Stepinski

Robert E Rhoads

Edward Darzynkiewicz

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 P-3'-deoxy-7-methylguanosine-5&#x2032;P-guanosine-5&#x2032;triphosphate and P-3&#x2032;-O,7-dimethylguanosine-5&#x2032;P-guanosine-5&#x2032;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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Warsaw, Poland

Janusz Stepinski