Diploid transgenic organisms are either hemi- or homozygous. Genetic assays are, therefore, required to identify the genotype. Our AGameOfClones vector concept uses two clearly distinguishable transformation markers embedded in interweaved, but incompatible Lox site pairs. Cre-mediated recombination leads to hemizygous individuals that carry only one marker. In the following generation, heterozygous descendants are identified by the presence of both markers and produce homozygous progeny that are selected by the lack of one marker. We prove our concept in Tribolium castaneum by systematically creating multiple functional homozygous transgenic lines suitable for long-term fluorescence live imaging. Our approach saves resources and simplifies transgenic organism handling. Since the concept relies on the universal Cre-Lox system, it is expected to work in all diploid model organisms, for example, insects, zebrafish, rodents and plants. With appropriate adaptions, it can be used in knock-out assays to preselect homozygous individuals and thus minimize the number of wasted animals.

中文翻译：

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用于转基因生物直接基因分型和系统创建纯合系的通用载体概念

二倍体转基因生物是半合子或纯合子。因此，需要进行基因检测来确定基因型。我们的 AGameOfClones 向量概念使用嵌入在交织但不兼容的 Lox 位点对中的两个清晰可辨的转换标记。Cre 介导的重组导致只携带一个标记的半合子个体。在下一代中，杂合子代通过两种标记的存在来鉴定，并产生通过缺少一种标记而选择的纯合子代。我们通过系统地创建适用于长期荧光实时成像的多种功能纯合转基因品系，证明了我们在 Tribolium castaneum 中的概念。我们的方法节省了资源并简化了转基因生物的处理。由于该概念依赖于通用 Cre-Lox 系统，预计它适用于所有二倍体模式生物，例如昆虫、斑马鱼、啮齿动物和植物。通过适当的调整，它可以用于敲除试验以预选纯合个体，从而最大限度地减少浪费动物的数量。