Manipulation of mouse embryonic stem cells for knockout mouse production.
Manipulation of mouse embryonic stem cells for knockout mouse production.
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DOI：
10.1002/0471143030.cb1913s44
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发表时间：
2009-09
影响因子：
--
通讯作者：
Kulkarni, Ashok B
中科院分区：
文献类型：
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作者：
Limaye, Advait;Hall, Bradford;Kulkarni, Ashok B
小鼠胚胎干（ES）细胞系的建立为基因敲除小鼠的产生提供了可能。然后可以操作在培养物中遗传改变的ES细胞以获得含有所需突变的完整小鼠。然而，为了成功地产生基因敲除小鼠，在整个基因靶向实验中，ES细胞必须以多能状态小心地培养。本单元详细介绍了使用同源重组技术成功产生基因敲除胚胎干细胞所需的一步一步的协议，试剂，设备和策略。
The establishment of mouse embryonic stem (ES) cell liness has allowed for the generation of the knockout mouse. ES cells that are genetically altered in culture can then be manipulated to derive a whole mouse containing the desired mutation. To successfully generate a knockout mouse, however, the ES cells must be carefully cultivated in a pluripotent state throughout the gene targeting experiment. This unit describes detailed step-by-step protocols, reagents, equipment, and strategies needed for the successful generation of gene knockout embryonic stem cells using homologous recombination technologies.