Endocrinology/Diabetes and Bone Disease Division, Mount Sinai School of Medicine, New York, New York, United States of America.
PLoS One. 2009 Nov 26;4(11):e8054. doi: 10.1371/journal.pone.0008054.
Conditional gene manipulation in mice becomes a routine for genetic studies of mammalian gene functions. Additional site-specific recombinases such as FLP or phi31 provide one more level of gene manipulation flexibility. The recombination activity of the currently available FLP deleter mice remains low. We generated a new FLP deleter mouse line with the mouse codon-optimized FLPo gene in C57BJ/6 background, which showed superior recombination efficacy in comparison to FLPe deleter mice. 100% complete removal of FRT-flanked Neo cassette was observed in all F1 progeny mice carrying both FLPo and Neo cassette, which can be transmitted to F2 generation independent of FLPo activity. Our new FLPo transgenic mice (on pure C57BJ/6 background) will largely facilitate the gene targeting process and is valuable for conditional gene manipulation.
条件性基因操作在小鼠中已成为研究哺乳动物基因功能的常规手段。额外的位点特异性重组酶,如 FLP 或 phi31,为基因操作提供了更多的灵活性。目前可用的 FLP 缺失小鼠的重组活性仍然较低。我们在 C57BJ/6 背景下生成了一种新型的 FLP 缺失小鼠系,该小鼠系携带经过密码子优化的鼠源 FLPo 基因,与 FLPe 缺失小鼠相比,其重组活性更高。在携带 FLPo 和 Neo 盒的所有 F1 后代小鼠中,均观察到 FRT 侧翼的 Neo 盒完全缺失,该缺失可以不依赖于 FLPo 的活性传递到 F2 代。我们的新型 FLPo 转基因小鼠(纯 C57BJ/6 背景)将极大地促进基因靶向过程,对于条件性基因操作非常有价值。