Optimization of insect odorant receptor trafficking and functional expression 
via transient transfection in HEK293 cells

Miazzi, Fabio; Hoyer, Carolin; Sachse, Silke; Knaden, Markus; Wicher, Dieter; Hansson, Bill S.; Lavista Llanos, Sofia

doi:10.1093/chemse/bjz062

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Optimization of insect odorant receptor trafficking and functional expression via transient transfection in HEK293 cells

MPS-Authors

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Miazzi, Fabio
Department of Evolutionary Neuroethology, Prof. B. S. Hansson, MPI for Chemical Ecology, Max Planck Society;
IMPRS on Ecological Interactions, MPI for Chemical Ecology, Max Planck Society;

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Hoyer, Carolin
Department of Evolutionary Neuroethology, Prof. B. S. Hansson, MPI for Chemical Ecology, Max Planck Society;

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Sachse, Silke
Research Group Dr. S. Sachse, Olfactory Coding, MPI for Chemical Ecology, Max Planck Society;

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Knaden, Markus
Research Group Dr. M. Knaden, Insect Behavior, Department of Neuroethology, Prof. B. S. Hansson, MPI for Chemical Ecology, Max Planck Society;

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Wicher, Dieter
Department of Evolutionary Neuroethology, Prof. B. S. Hansson, MPI for Chemical Ecology, Max Planck Society;

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Hansson, Bill S.
Department of Evolutionary Neuroethology, Prof. B. S. Hansson, MPI for Chemical Ecology, Max Planck Society;

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Lavista Llanos, Sofia
Research Group Dr. M. Stensmyr, Molecular Biology of Olfaction, Department of Neuroethology, Prof. B. S. Hansson, MPI for Chemical Ecology, Max Planck Society;

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http://dx.doi.org/10.1093/chemse/bjz062
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HAN335.pdf
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Supplementary Material (public)

HAN335s1.pdf
(Supplementary material), 2MB

Citation

Miazzi, F., Hoyer, C., Sachse, S., Knaden, M., Wicher, D., Hansson, B. S., et al. (2019). Optimization of insect odorant receptor trafficking and functional expression via transient transfection in HEK293 cells. Chemical Senses, 44(9), 673-682. doi:10.1093/chemse/bjz062.

Cite as: https://hdl.handle.net/21.11116/0000-0003-CBC9-6

Abstract

Insect odorant receptors show a limited functional expression in various heterologous expression systems including insect and mammalian cells. This may be in part due to the absence of key components driving the release of these proteins from the endoplasmic reticulum and directing them to the plasma membrane. In order to mitigate this problem we took advantage of small export signals within the human HCN1 and Rhodopsin that have been shown to promote protein release from the endoplasmic reticulum and the trafficking of post-Golgi vesicles, respectively. Moreover, we designed a new vector based on a bidirectional expression cassette to drive the functional expression of the insect odorant receptor co-receptor (Orco) and an odor-binding odorant receptor, simultaneously. We show that this new method can be used to reliably express insect odorant receptors in HEK293 cells via transient transfection and that is highly suitable for downstream applications using automated and high-throughput imaging platforms.