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Abstract

The invention relates to a method for the production of a eukaryotic cell selectable by inactivation or reduction of an endogenous gene function, comprising the steps of (a) introduction of one or more vectors into the cell and (b) expression of a siRNA and preferably shRNA coded by the one or more vectors, directed against an endogenous selectable gene and inactivating same, said siRNA or shRNA being the transcription product of an RNAi selection cassette, the selection cassette comprising a section of at least 19 nucleotides of the transcribed region of the gene, said selection being operatively linked to a promoter and a transcription termination signal. Furthermore, the invention relates to a eukaryotic cell comprising an RNAi selection cassette which is directed against an endogenous selectable gene and inactivates the function of said gene; wherein the RNAi selection cassette comprises a section of the gene with a length of at least 19 nucleotides which is operatively linked to a promoter and a transcription termination signal. Moreover, the invention relates to methods for the production of a transgenic mammal, comprising the steps: (a) injection of the embryonic stem cell according to the invention or of an embryonic stem cell selected according to the method of the invention in blastocysts of a mammal, (b) transfer of the blastocysts into the uterus of a mammal, and (c) carrying the transgenic mammal to full term. Finally, the invention relates to a method for the production of a transgenic plant.

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First Claim