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T7-O-me-RNA-Polymerase Tailored Polymerase for efficient synthesis of stable RNA Invention RNA is increasingly being used as a tool in a broad range of applications, e.g. as aptamers, in particular as an active agent in innovative therapeutic approaches. However, these applications require the modification of RNA which otherwise shows poor resistance to cellular nucleases. To this end, typically the ribose 2´hydroxyl moieties are substituted by O-methyl groups. This step is hampered by the fact that wild-type RNA-Polymerase is inefficient in incorporating modified nucleotides. Although mutant enzymes have been engineered which are able to insert some or even all 2´-O-me-modified nucleotides, none of the enzymes present so far is able to synthesize transcripts of sufficient lengths. Ribose-specific interactions between nucleotides and By an in-depth analysis of the T7active site residues of wild-type T7-RNA-Polymerase RNA-Polymerase and protein engineering, the inventors were able to create a variant which is both a generalist polymerase capable of incorporating all 2´-O-me-modified nucleotides as well as synthesizing fully modified RNA of up to approx. 1,000 nucleotides. In vitro transcription of 284-nt template using inventive Competitive Advantages Long-range synthesis of 2´-Ome-RNA for obtaining stable RNA Faithful incorporation of all 2´-O-me-RNA-nucleotides Robust enzyme for a broad range of applications including easy & efficient synthesis of aptamers enzyme and 2´-O-me-modified NTP as indicated In vitro transcription of 1,000-nt template using inventive enzyme. r: all NTPs; other lanes: only indicated NTP modified; all: all NTPs modified Commercial Opportunities The inventive enzyme is broadly applicable in molecular and cellular biology both in academia and in industry. Furthermore, the inventive enzyme is a key tool for developing novel RNAbased therapeutic approaches. Current Status The inventive polymerase thereof has been tested in numerous laboratory applications and has proven to be a robust tool. On behalf of the Technical University Dortmund, PROvendis offers the invention for licensing. A priority patent application has been filed. Relevant Publication Ibach, J. et al. (2013) “Identification of a T7 RNA polymerase variant that permits the enzymatic synthesis of fully 2'-O-methyl-modified RNA”. J. Biotechnol. 167(3): 287-9. Kontakt: Ref.-Nr.: 2768 Dr. Juergen Walkenhorst PROvendis GmbH is the patent licensing agency for the universities of North Rhine-Westphalia, Germany. PROvendis recommends: www.inventionstore.de - Free e-mail service to access the latest IP-protected top technologies. PROvendis GmbH Schloßstraße 11-15 D-45468 Mülheim an der Ruhr Tel.: +49 (0)208 94 105 25 Fax: +49 (0)208 94 105 50 E-Mail: [email protected] Web: www.provendis.info