A call for caution in analysing mammalian co-transfection experiments and implications of resource competition in data misinterpretation

Roberto Di Blasi, Masue M. Marbiah, Velia Siciliano, Karen Polizzi and Francesca Ceroni ()
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Roberto Di Blasi: Imperial College London, South Kensington Campus
Masue M. Marbiah: Imperial College London, South Kensington Campus
Velia Siciliano: Istituto Italiano di Tecnologia-IIT, Largo Barsanti e Matteucci
Karen Polizzi: Imperial College London, South Kensington Campus
Francesca Ceroni: Imperial College London, South Kensington Campus

Nature Communications, 2021, vol. 12, issue 1, 1-6

Abstract: 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.

Date: 2021
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DOI: 10.1038/s41467-021-22795-9

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