Icon Genetics Gmbh

Icon Genetics Ag

Carola Engler

Sylvestre Marillonnet

Yuri Gleba

Victor Klimyuk

Stefan Werner

Ernst Weber

Romy Kandzia

Frank Thieme

Stefan Mühlbauer

Stefan Herz

Ramona Grutzner

Stefan Muehlbauer

Ramona Gruetzner

System for producing a nucleic acid construct of interest, said system comprising:

System and method of modular cloning

A process of expressing a sequence of interest in a plant, plant part, or plant cell culture, comprising: (a) providing a plant, plant part, or plant cell culture containing in cell nuclei a heterologous DNA having a sequence encoding an RNA replicon operably linked or linkable to a transcription promoter, wherein said sequence encoding an RNA replicon contains (i) sequences for replicon function of said RNA replicon, said sequences being derived from a sequence of a plant RNA virus, (ii) a sequence of interest, whereby said sequences for replicon function exhibit at selected localities of said sequences of said plant RNA virus function-conservative differences from said sequence of said plant RNA virus, said differences causing an increased frequency of replicon formation compared to an RNA replicon not exhibiting said differences; and (b) causing expression of said sequence of interest.

RNA virus-derived plant expression system

A process of expressing a sequence of interest in a plant, plant part, or plant cell culture, comprising: (a) providing a plant, plant part, or plant cell culture containing in cell nuclei a heterologous DNA having a sequence encoding an RNA replicon operably linked or linkable to a transcription promoter, wherein said sequence encoding an RNA replicon contains (i) sequences for replicon function of said RNA replicon, said sequences being derived from a sequence of a plant RNA virus, (ii) a sequence of interest, whereby said sequences for replicon function exhibit at selected localities of said sequences of said plant RNA virus function conservative differences from said sequence of said plant RNA virus, said differences causing an increased frequency of replicon formation compared to an RNA replicon not exhibiting said differences; and (b) causing expression of said sequence of interest.

A process of inserting a nucleic acid sequence of interest into an acceptor nucleic acid is provided. The process comprises amplifying by PCR a DNA comprising in the following order a sequence segment U, a nucleic acid sequence segment of known nucleotide sequence K2, and a nucleic acid sequence segment of known sequence K3. The process further comprises treating the linear double-stranded DNA molecules from the PCR amplification with an exonuclease to obtain a single-stranded overhang at the first end of the DNA and a single-stranded overhang comprising nucleic acid segments K2 and K3 at the second end of the DNA. The process additionally comprises annealing the product of the exonuclease treatment to a linearized double-stranded acceptor nucleic acid which has been designed to complement the single-stranded overhangs of the product of the exonuclease treatment.

Process of clean cloning

A process for replicating or for replicating and expressing a sequence of interest in a plant, comprising: (i) an RNA replicon or a precursor thereof, said RNA replicon being derived from a plus-sense single stranded RNA virus and comprising at least one sequence of interest; and (ii) a helper replicon, or a precursor thereof, wherein said helper replicon is (a) incapable of systemic movement in said plant both in the presence and in the absence of said RNA replicon (i) and (b) capable of expressing in a plant one or more proteins necessary for systemic movement of said RNA replicon (i), whereby said RNA replicon (i) is capable of replicating or replicating and expressing said sequence of interest in said plant, but unable to move systemically in said plant in the absence of said one or more proteins expressed by said helper replicon (ii).

Two-component RNA virus-derived plant expression system

Nucleic acid comprising or encoding an RNA replica comprising, in this order, the following segments (i) to (iii): i) a nucleic acid sequence encoding a potexvirus RNA-dependent RNA polymerase or a function-conservative variant thereof; ii) a nucleic acid sequence comprising: a) a potexvirus triple gene block or a function-conservative variant thereof and b) a sequence encoding a potexviral coat protein or a function-conservative variant thereof; or a sequence encoding a tobago viral movement protein; and iii) a heterologous nucleic acid sequence expressible from said replica in a plant or in plant tissue.

Potexvirus-derived replicon

A process of producing a protein of interest by expression of said protein of interest from a sequence of interest in a plant or in plant leaves, comprising: (a) transfecting said plant or said plant leaves by infiltrating said plant or said plant leaves with anstrain in the presence of a complementing factor, saidstrain containing in T-DNA a heterologous DNA sequence having a sequence portion encoding a replicon, wherein said sequence encoding a replicon contains sequences necessary for replicon function of said replicon, said sequences being derived from a plant virus, and said sequence of interest to be expressed from said replicon, (b) optionally isolating said protein of interest from said plant or said plant leaves infiltrated in step (a), wherein saidstrain is provided with a first genetic modification rendering saidstrain defective for transfecting organisms with said T-DNA in the absence of said complementing factor.

Growing community of inventors

Halle, Germany

Carola Engler