The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Sep. 04, 2012

Filed:

Sep. 03, 2002
Applicants:

Joseph Atabekov, Moscow, RU;

Yurii Dorokhov, Moscow, RU;

Maxim Skulachev, Moscow, RU;

Peter Ivanov, Moscow, RU;

Yuri Gleba, München, DE;

Inventors:

Joseph Atabekov, Moscow, RU;

Yurii Dorokhov, Moscow, RU;

Maxim Skulachev, Moscow, RU;

Peter Ivanov, Moscow, RU;

Yuri Gleba, München, DE;

Assignee:

Icon Genetics, Inc., Princeton, NJ (US);

Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12N 15/00 (2006.01); C12N 15/09 (2006.01); C12N 15/11 (2006.01); C12N 15/63 (2006.01); C12N 15/67 (2006.01); C12N 15/82 (2006.01); C12N 15/85 (2006.01);
U.S. Cl.
CPC ...
Abstract

A method of search for and identification of a eukaryotic IRES element active in cap-independent translation of RNA in eukaryotic cells is provided, comprising the following steps: i) screening eukaryotic mRNA sequences or corresponding DNA sequences for a potential IRES element having a block of nucleotides having: a) a length of at least 30 nucleotides; b) an adenine nucleotide content of at least 40 mol-%; and c) a pyrimidine nucleotide content of less than 40 mol-%; ii) inserting said potential IRES element into a linear dicistronic construct between an upstream gene and a downstream GUS reporter gene, whereby said potential IRES element is positioned for IRES-dependent translation of said downstream GUS gene and whereby said upstream gene is preceded by a stable hairpin structure to prevent IRES-independent translation of said genes; and iii) testing said potential IRES element for IRES-dependent translation of said GUS gene in a rabbit reticulocyte lysate or in a wheat germ extract in vitro translation assay, whereby GUS gene expression is quantitated preferably relative to a construct having a reference IRES element or a non-IRES element between said upstream gene and said GUS gene.


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