Synthesis and use of anti-reverse mRNA cap analogues
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United States of America Patent
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Jul 11, 2006
Grant Date -
Oct 16, 2003
app pub date -
May 17, 2002
filing date -
Mar 25, 2002
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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 P.sup.1-3'-deoxy-7-methylguanosine-5'P.sup.3-guanosine-5'triphosphate and P.sup.1-3'-O,7-dimethylguanosine-5'P.sup.3-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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