Simultaneous generation of fra-2 conditional and fra-2 knock-out mice
Simultaneous generation of fra-2 conditional and fra-2 knock-out mice
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DOI：
10.1002/dvg.20311
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发表时间：
2007-07-01
期刊：
影响因子：
1.5
通讯作者：
Wagner, Erwin F.
中科院分区：
文献类型：
--
作者：
Eferl, Robert;Zenz, Rainer;Wagner, Erwin F.
功能丧失的小鼠模型包括生殖系中基因缺失的基因敲除小鼠，以及特定组织中存在体细胞等位基因缺失的条件性基因敲除小鼠。这两种类型的小鼠都用于体内基因功能的全面研究。在这里，我们描述了一种简单的方法，使用单个胚胎干细胞克隆同时产生转录因子fra-2(Fos相关抗原2)的条件等位基因或敲除等位基因的小鼠。将含有FRA-2等位基因的ES细胞瞬时导入Cre重组酶表达载体，低密度平板培养。大多数由此产生的ES细胞克隆由保留或丢失条件等位基因的细胞的混合物组成。我们证明，这些混合的ES细胞克隆可以直接用于产生嵌合体，这些嵌合体同时产生带有条件等位基因或敲除等位基因的后代。这一策略缩短了产生转基因小鼠的时间并减少了生殖系传播事件的数量。
Loss of function mouse models comprise knock-out mice, where a gene is deleted in the germline, and conditional knock-out mice with somatic deletion of a floxed allele in defined tissues. Both types of mice are used for comprehensive studies of gene functions in vivo. Here, we describe a simple method for simultaneous generation of mice with conditional or knock-out alleles for the transcription factor fra-2 (Fos-related antigen 2) using a single embryonic stem (ES) cell clone. ES cells with a floxed fra-2 allele were transiently transfected with a Cre-recombinase expression plasmid and plated at low density. Most of the resulting ES cell colonies consisted of a mixture of cells that have either retained or lost the conditional allele. We demonstrate that these mixed ES cell clones can be directly used for generation of chimeras that give rise to offspring with conditional or knock-out alleles simultaneously. This strategy shortens the time and reduces the number of germline transmission events to generate genetically modified mice.