The doxycycline inducible overexpression system is a highly flexible and widely used tool for both in vitro and in vivo studies. However, during the past decade, a handful of reports have explicitly called for caution when using this system. The raised concerns are based on the notion that doxycycline can impair mitochondrial function of mammalian cells and can alter properties such as cell proliferation. As such, experimental outcomes can be confounded with the side effects of doxycycline and valid interpretation can be seriously threatened. Today, no consensus seems to exist about how these problems should be prevented. Moreover, some of the strategies that have been used to cope with these difficulties can actually introduce additional problems that are related to genomic instability and genetic modification of the cells. Here, we elaborate on the above statements and clarify them by some basic examples taken from our personal wet-lab experience. As such, we provide a nuanced overview of the doxycycline inducible overexpression system, some of its limitations and how to deal with them.

强力霉素诱导过表达系统是一种高度灵活且广泛用于体外和体内研究的工具。然而，在过去十年中，一些报告明确呼吁在使用该系统时要谨慎。提出的担忧是基于多西环素会损害哺乳动物细胞的线粒体功能并改变细胞增殖等特性的观点。因此，实验结果可能与多西环素的副作用混淆，有效解释可能会受到严重威胁。今天，似乎没有就如何预防这些问题达成共识。此外，用于应对这些困难的一些策略实际上可能会引入与基因组不稳定性和细胞遗传修饰相关的其他问题。这里，我们详细阐述了上述陈述，并通过我们个人湿实验室经验中的一些基本例子来阐明它们。因此，我们对强力霉素诱导的过度表达系统、它的一些局限性以及如何处理它们进行了细致入微的概述。