FO generation mice fully derived from gene-targeted embryonic stem cells allowing immediate phenotypic analyses
FO generation mice fully derived from gene-targeted embryonic stem cells allowing immediate phenotypic analyses
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DOI：
10.1038/nbt1263
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发表时间：
2007-01-01
影响因子：
46.9
通讯作者：
Valenzuela, David M.
中科院分区：
文献类型：
--
作者：
Poueymirou, William T.;Auerbach, Wojtek;Valenzuela, David M.
探索基因功能的一种有用的方法是从转基因的胚胎干细胞产生突变小鼠。胚胎干细胞基因工程的最新进展将这一过程中的瓶颈转移到了小鼠的生成上。传统的胚胎干细胞注射到囊胚宿主产生的F0代嵌合体只有一部分来自胚胎干细胞，需要额外的育种来获得可以表型的突变小鼠。四倍体互补方法直接产生几乎完全来自ES细胞的小鼠，但它效率低下，仅适用于某些杂交ES细胞系，并患有非特异性致死性和异常，使表型分析复杂化。在这里，我们展示了激光辅助将近交系或杂交ES细胞注射到8个细胞期胚胎中，有效地产生了完全由ES细胞衍生的健康F0代小鼠，表现出100%的生殖系传播率，并允许立即进行表型分析，极大地加快了基因功能分配。
A useful approach for exploring gene function involves generating mutant mice from genetically modified embryonic stem (ES) cells. Recent advances in genetic engineering of ES cells have shifted the bottleneck in this process to the generation of mice. Conventional injections of ES cells into blastocyst hosts produce F0 generation chimeras that are only partially derived from ES cells, requiring additional breeding to obtain mutant mice that can be phenotyped. The tetraploid complementation approach directly yields mice that are almost entirely derived from ES cells, but it is inefficient, works only with certain hybrid ES cell lines and suffers from nonspecific lethality and abnormalities, complicating phenotypic analyses. Here we show that laser-assisted injection of either inbred or hybrid ES cells into eight cell-stage embryos efficiently yields F0 generation mice that are fully ES cell-derived and healthy, exhibit 100% germline transmission and allow immediate phenotypic analysis, greatly accelerating gene function assignment.