Download T7-O-me-RNA-Polymerase

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