Optimization of the Doxycycline-Induced Gene Expression System in HCT116 and A549 Cells

被引：0
作者：
A. M. Kozlova [1 ]
A. V. Morshneva [1 ]
O. O. Gnedina [1 ]
M. V. Igotti [1 ]
机构：
[1] Institute of Cytology, Russian Academy of Sciences, St. Petersburg
来源：
Cell and Tissue Biology | 2025年 / 19卷 / 2期
基金：
俄罗斯科学基金会;
关键词：
cell viability; doxycycline; inducible expression of gene of interest; proliferation;
D O I：
10.1134/S1990519X24600777
中图分类号：
学科分类号：
摘要：
Abstract: Objective: The doxycycline (Dox)-inducible gene expression system, which allows for the overexpression of a gene of interest under the control of the Tet-operator, is a practical and widely used approach in both in vitro and in vivo studies. However, recent observations indicate the possibility of antiproliferative and/or cytotoxic effects of doxycycline in certain cell systems. Consequently, the results of studies using Dox-inducible expression systems may be misinterpreted or confounded by the side effects of doxycycline itself. Methods and Results: In this study, we determined the lowest concentration of doxycycline required for efficient induction of the gene of interest under the control of the Tet-operator and analyzed its impact on the proliferation and viability of two human tumor cell lines HCT116 and A549, which are widely used in scientific research. Conclusions: Our findings highlight the importance of including additional controls in experiments using Dox-inducible expression systems. Specifically, the effects of the applied doxycycline concentrations should be assessed in cells that do not carry the Dox-inducible gene. This step is critical to avoid artifacts arising from the direct effects of doxycycline on the studied parameters. © Pleiades Publishing, Ltd. 2025.
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页码：151 / 160
页数：9
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