A call for caution in analysing mammalian co-transfection experiments and implications of resource competition in data misinterpretation.
A call for caution in analysing mammalian co-transfection experiments and implications of resource competition in data misinterpretation.
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DOI：
10.1038/s41467-021-22795-9
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发表时间：
2021-05-05
影响因子：
16.6
通讯作者：
Ceroni F
中科院分区：
文献类型：
--
作者：
Di Blasi R;Marbiah MM;Siciliano V;Polizzi K;Ceroni F
瞬时转染通常用于基础和合成生物学研究，以解开通路调节和探针和探针电路设计。由于每个实验都有内在变异性的成分，报告基因表达通常用作为转染对照的共递送基因进行标准化。最近在哺乳动物细胞中的报告强调了基因表达的资源竞争如何导致数据解释的偏差，对共转染实验产生直接影响。在这里，我们定义资源竞争和瞬时转染实验之间的连接，并讨论可能的替代方案。我们的目标是提高社区内的认识，并激发讨论，将这些考虑因素纳入未来的实验设计中，以开发更好的转染控制。
Transient transfections are routinely used in basic and synthetic biology studies to unravel pathway regulation and to probe and characterise circuit designs. As each experiment has a component of intrinsic variability, reporter gene expression is usually normalized with co-delivered genes that act as transfection controls. Recent reports in mammalian cells highlight how resource competition for gene expression leads to biases in data interpretation, with a direct impact on co-transfection experiments. Here we define the connection between resource competition and transient transfection experiments and discuss possible alternatives. Our aim is to raise awareness within the community and stimulate discussion to include such considerations in future experimental designs, for the development of better transfection controls.
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