公开(公告)号：US20110041196A1

公开(公告)日：2011-02-17

申请号：US12856126

申请日：2010-08-13

申请人： David FRENDEWEY ,  David Jonathan HESLIN ,  Ka-Man Venus LAI ,  David M. VALENZUELA

发明人： David FRENDEWEY ,  David Jonathan HESLIN ,  Ka-Man Venus LAI ,  David M. VALENZUELA

IPC分类号： C12N15/87 ,  C12N15/85 ,  C12N5/10

CPC分类号： C12N15/85 ,  A01K67/0275 ,  A01K67/0276 ,  A01K2217/07 ,  A01K2217/075 ,  A01K2217/206 ,  A01K2227/105 ,  A01K2267/03 ,  C12N5/0606 ,  C12N9/1241 ,  C12N15/8509 ,  C12N15/873 ,  C12N15/907 ,  C12N2800/22 ,  C12N2800/30 ,  C12N2840/102 ,  C12Y207/07

摘要： Targeting constructs and methods of using them are provided for differentiation-dependent modification of nucleic acid sequences in cells and in non-human animals. Targeting constructs comprising a promoter operably linked to a recombinase are provided, wherein the promoter drives transcription of the recombinase in an differentiated cell but not an undifferentiated cell. Promoters include Blimp1, Prm1, Gata6, Gata4, Igf2, Lhx2, Lhx5, and Pax3. Targeting constructs with a cassette flanked on both sides by recombinase sites can be removed using a recombinase gene operably linked to a 3′-UTR that comprises a recognition site for an miRNA that is transcribed in undifferentiated cells but not in differentiated cells. The constructs may be included in targeting vectors, and can be used to automatically modify or excise a selection cassette from an ES cell, a non-human embryo, or a non-human animal.

摘要翻译： 提供靶向构建体和使用它们的方法用于细胞和非人动物中核酸序列的分化依赖性修饰。 提供了包含与重组酶可操作地连接的启动子的靶向构建体，其中所述启动子驱动分化细胞中但不是未分化细胞的重组酶的转录。 启动子包括Blimp1，Prm1，Gata6，Gata4，Igf2，Lhx2，Lhx5和Pax3。 可以使用可操作地连接到3'-UTR的重组酶基因来除去具有通过重组酶位点两侧两侧的盒的靶向构建体，其包含在未分化细胞中但不在分化细胞中转录的miRNA的识别位点。 构建体可以包括在靶向载体中，并且可以用于自动修饰或从ES细胞，非人胚胎或非人动物中切除选择盒。

公开(公告)号：US20090271884A1

公开(公告)日：2009-10-29

申请号：US12399109

申请日：2009-03-06

申请人： William POUEYMIROU ,  Thomas M. DECHIARA ,  Wojtek AUERBACH ,  Aris N. ECONOMIDES ,  Nicholas W. GALE ,  David FRENDEWEY ,  David M. VALENZUELA

发明人： William POUEYMIROU ,  Thomas M. DECHIARA ,  Wojtek AUERBACH ,  Aris N. ECONOMIDES ,  Nicholas W. GALE ,  David FRENDEWEY ,  David M. VALENZUELA

IPC分类号： C12N15/85 ,  C12N5/06

CPC分类号： A01K67/0271 ,  A01K67/0275 ,  A01K2207/12 ,  A01K2217/00 ,  A01K2217/05 ,  A01K2217/07 ,  A01K2217/15 ,  A01K2217/203 ,  A01K2217/30 ,  A01K2227/105 ,  A01K2267/02 ,  C12N5/16 ,  C12N15/8509 ,  C12N15/907 ,  C12N2510/00 ,  C12N2517/02

摘要： Genetically modified mice and nucleic acid constructs for making the genetically modified mice are described. A first mouse having a gene encoding an activator (such as a Cre recombinase) operably linked to a developmentally-regulated promoter (such as a Nanog promoter) is provided. A second mouse having a toxic responder gene (such as a gene encoding diphtheria toxin A) is provided, where the toxic gene is expressed only in the presence of an activator, Embryos from a mating of the first and the second mouse are provided as host embryos suitable for generating mice from donor cells introduced into the host embryos. Ablating the ICM of a mouse embryo physically, chemically, or genetically is described, as well as making F0 generation mice that are substantially or in full derived from donor cells, employing a host mouse embryo with an ablated or nonproliferating ICM.

摘要翻译： 描述了用于制备遗传修饰的小鼠的转基因小鼠和核酸构建体。 提供了具有编码与发育调节的启动子（例如Nanog启动子）可操作地连接的活化剂（例如Cre重组酶）的基因的第一只小鼠。 提供具有毒性反应基因（例如编码白喉毒素A的基因）的第二只小鼠，其中毒性基因仅在活化剂存在下表达，来自第一和第二小鼠的交配的胚胎被提供为宿主 适用于从供体细胞产生小鼠的胚胎引入宿主胚胎。 描述了在物理，化学或遗传学上消除小鼠胚胎的ICM，以及使用具有消融或非增殖ICM的宿主小鼠胚胎制备基本上或完全衍生自供体细胞的F0代小鼠。