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

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.

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 Creators:
Miazzi, Fabio1, 2, Author           
Hoyer, Carolin1, Author           
Sachse, Silke3, Author           
Knaden, Markus4, Author           
Wicher, Dieter1, Author           
Hansson, Bill S.1, Author           
Lavista Llanos, Sofia5, Author           
Affiliations:
1Department of Evolutionary Neuroethology, Prof. B. S. Hansson, MPI for Chemical Ecology, Max Planck Society, ou_421894              
2IMPRS on Ecological Interactions, MPI for Chemical Ecology, Max Planck Society, Jena, DE, ou_421900              
3Research Group Dr. S. Sachse, Olfactory Coding, MPI for Chemical Ecology, Max Planck Society, ou_421914              
4Research Group Dr. M. Knaden, Insect Behavior, Department of Neuroethology, Prof. B. S. Hansson, MPI for Chemical Ecology, Max Planck Society, ou_421913              
5Research Group Dr. M. Stensmyr, Molecular Biology of Olfaction, Department of Neuroethology, Prof. B. S. Hansson, MPI for Chemical Ecology, Max Planck Society, ou_421915              

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 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.

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 Dates: 2019-08-3120192019-10-26
 Publication Status: Issued
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 Identifiers: Other: HAN335
DOI: 10.1093/chemse/bjz062
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Title: Chemical Senses
Source Genre: Journal
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Publ. Info: Oxford : Oxford University Press [etc.]
Pages: - Volume / Issue: 44 (9) Sequence Number: - Start / End Page: 673 - 682 Identifier: ISSN: 0379-864X
CoNE: https://pure.mpg.de/cone/journals/resource/954928560444