Synthesis and use of anti-reverse mRNA cap analogues

US7074596B2 · kind B2 · utility

Assignee

• BOARD OF SUPERVISORS OF LOUISIANA STATE UNIVERSITY AND AGRICULTURAL AND MECHANICAL COLLEGE · US

Inventors

• Edward Darzynkiewicz · Białystok, PL
• Robert E. Rhoads · Shreveport, US
• Janusz Stepinski · Warszawa, PL

Key dates

• Filing date: May 17, 2002
• Grant date: Jul 11, 2006
• Expiry date: Jun 20, 2023

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12P21/02
• WIPO field: Biotechnology
• WIPO sector: Chemistry

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.