Kranz Andrea, Fu Jun, Duerschke Kristin, Weidlich Stefanie, Naumann Ronald, Stewart A Francis, Anastassiadis Konstantinos
Genomics, BioInnovationsZentrum, Technische Universität Dresden, Am Tatzberg 47, Dresden, Germany.
Genesis. 2010 Aug;48(8):512-20. doi: 10.1002/dvg.20641.
Recently, a codon improved version of the Flpe site specific recombinase, termed Flpo, was reported as having greatly improved performance in mammalian cell applications. However, the degree of improvement could not be estimated because essentially no Flpe activity was observed. Here, we compare Flpe and Flpo accurately in a mammalian cell assay to estimate that Flpo is about five times more active than Flpe and similar to Cre and Dre. Consequently, we generated a Flpo deleter mouse line from the JM8 C57Bl/6 ES cells used in the EUCOMM and KOMP systematic knock-out programs. In breeding experiments, we show that the Flpo deleter delivers complete recombination using alleles that are incompletely recombined by a commonly used Flpe deleter. This indicates that the Flpo deleter is more efficient.
最近,据报道,一种密码子优化的Flpe位点特异性重组酶版本,称为Flpo,在哺乳动物细胞应用中具有显著提高的性能。然而,由于基本上未观察到Flpe活性,因此无法估计其改善程度。在这里,我们在哺乳动物细胞试验中准确比较了Flpe和Flpo,估计Flpo的活性约为Flpe的五倍,与Cre和Dre相似。因此,我们从欧盟共同执行体计划(EUCOMM)和敲除小鼠计划(KOMP)系统敲除程序中使用的JM8 C57Bl/6胚胎干细胞(ES细胞)中产生了一个Flpo删除小鼠品系。在繁殖实验中,我们表明,Flpo删除品系使用常用Flpe删除品系不完全重组的等位基因实现了完全重组。这表明Flpo删除品系效率更高。