Icon Genetics Gmbh

Yurii Dorokhov

Yuri Gleba

Peter Ivanov

Joseph Atabekov

Maxim Vladimirovich Skulachev

Victor Klimyuk

Sylvestre Marillonnet

Stefan Werner

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.

Identification of eukaryotic internal ribosome entry site (IRES) elements

A method of producing a nucleic acid sequence having an adenine-rich nucleic acid block of at least 25 nucleotides in length is provided. The nucleic acid sequence that is produced is capable of initiating translation at an internal ribosome entry site (IRES) and the adenine-rich nucleic acid block comprises from 40 to 100 mol-% adenine.

Creation of artificial internal ribosome entry site (IRES) elements

A method of controlling a genetically-modified multi-cellular organism or a part thereof, comprising the following steps: (a) providing a multi-cellular organism or a part thereof, whereby cells of said multi-cellular organism or said part contain a heterologous nucleic acid, (b) causing expression of a I protein from said heterologous nucleic acid in at least some of said cells, wherein said protein is capable of (i) leaving a cell and entering other cells of said multi-cellular organism or a part thereof, (ii) causing expression of said protein in cells containing said heterologous nucleic acid, and optionally (iii) controlling a cellular process of interest.

Method of controlling a cellular process in a multi-cellular organism

The invention describes virus-based amplification vectors for plants containing additional plant-specific internal ribosome entry site (IRES) element(s) allowing for a polycistronic translation and a cap-independent translation of: a) heterologous gene(s); b) whole viral genome or c) viral subgenomic RNAs. Said IRES elements are of plant viral origin, or they are isolated from other organisms or engineered using different synthesis procedures. Said IRES element(s) and said heterologous gene(s) are inserted into amplification vectors and allow for the expression of said heterologous gene(s) in the absence of additional viral promoters, in particular, said expression is achieved through cap-independent translation.

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Moscow, Russia

Yurii Dorokhov