The power of Drosophila melanogaster as a model system relies on tractable germline genetic manipulations. Despite Drosophila’s expansive genetics toolbox, such manipulations are still accomplished one change at a time and depend predominantly on phenotypic screening. We describe a drug-based genetic platform consisting of four selection and two counterselection markers, eliminating the need to screen for modified progeny. These markers work reliably individually or in combination to produce specific genetic outcomes. We demonstrate three example applications of multiplexed drug-based genetics by generating (1) transgenic animals, expressing both components of binary overexpression systems in a single transgenesis step; (2) dual selectable and counterselectable balancer chromosomes; and (3) selectable, fluorescently tagged P[acman] bacterial artificial chromosome (BAC) strains. We perform immunoprecipitation followed by proteomic analysis on one tagged BAC line, demonstrating our platform’s applicability to biological discovery. Lastly, we provide a plasmid library resource to facilitate custom transgene design and technology transfer to other model systems.

果蝇作为模型系统的力量依赖于易于处理的种系遗传操作。尽管果蝇拥有广泛的遗传学工具箱，但此类操作仍然一次完成一个变化，并且主要依赖于表型筛选。我们描述了一个基于药物的遗传平台，由四个选择标记和两个反选择标记组成，消除了筛选修饰后代的需要。这些标记单独或组合可靠地发挥作用，产生特定的遗传结果。我们通过生成（1）转基因动物，在单个转基因步骤中表达二元过表达系统的两种成分，展示了基于多重药物的遗传学的三个示例应用； (2)双重选择性和反选择性平衡染色体； (3)可选择的、荧光标记的P[acman]细菌人工染色体(BAC)菌株。我们对一个标记的 BAC 系进行免疫沉淀，然后进行蛋白质组分析，证明我们的平台对生物发现的适用性。最后，我们提供质粒库资源，以促进定制转基因设计和技术转移到其他模型系统。