公开(公告)号：US20080120733A1

公开(公告)日：2008-05-22

申请号：US11629709

申请日：2005-06-16

申请人： Martin Hafner ,  Karina Nawrath ,  Werner Muller

发明人： Martin Hafner ,  Karina Nawrath ,  Werner Muller

IPC分类号： A01K67/027 ,  C12N15/87 ,  C12N5/02 ,  C12N5/00 ,  C12N5/04 ,  A01H1/00 ,  C12N15/00

CPC分类号： C12N15/8218 ,  C12N15/63 ,  C12N15/8209 ,  C12N15/821

摘要： 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.

摘要翻译： 本发明涉及通过灭活或还原内源基因功能来选择性地生产真核细胞的方法，其包括以下步骤：（a）将一种或多种载体导入细胞，和（b）表达siRNA，优选 由一个或多个载体编码的针对内源可选择基因并使其失活的shRNA，所述siRNA或shRNA是RNAi选择盒的转录产物，所述选择盒包含至少19个核苷酸的转录区的部分 基因，所述选择可操作地连接到启动子和转录终止信号。 此外，本发明涉及包含针对内源可选择基因并使所述基因的功能失活的RNAi选择盒的真核细胞; 其中所述RNAi选择盒包含长度为至少19个核苷酸的基因的一部分，其与启动子和转录终止信号有效连接。 此外，本发明涉及生产转基因哺乳动物的方法，其包括以下步骤：（a）根据本发明的胚胎干细胞或根据本发明的方法选择的胚胎干细胞在囊胚中的注射 哺乳动物，（b）将囊胚转移到哺乳动物的子宫中，和（c）将转基因哺乳动物全部携带。 最后，本发明涉及生产转基因植物的方法。