Odorant Receptor Choice Mechanism Revealed by Analysis of a Highly Represented Odorant Receptor Transgene

Makhlouf, Mélanie; D’Hulst, Charlotte; Omura, Masayo; Rosa, Alessandro; Mina, Raena; Bernal-Garcia, Sergio; Lempert, Eugene; Saraiva, Luís R.; Feinstein, Paul

doi:10.2139/ssrn.4119003

Cited by 2 publications

(2 citation statements)

References 32 publications

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“…S6, A and B). We previously characterized 5x21-OR1A1-Cherry (44) and showed that the dorsal epithelium expressed OR1A1 in 50% of the OSNs. Further increasing expression (OR abundance) in the dorsal epithelium to 100% of the OSNs may double the signal.…”

Section: Discussionmentioning

confidence: 99%

A genetic platform for functionally profiling odorant receptors in olfactory cilia ex vivo

et al. 2022

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The molecular basis for odor perception in humans remains enigmatic because of the difficulty in studying odorant receptors (ORs) outside their native environment. Efforts toward OR expression and functional profiling have been met with limited success because of the poor efficiency of their cell surface expression in vitro. Structures protruding from the surface of olfactory sensory neurons called cilia contain all of the components of the olfactory signal transduction machinery and can be placed in an ex vivo plate assay to rapidly measure odor-specific responses. Here, we describe an approach using cilia isolated from the olfactory sensory neurons of mice expressing two human ORs, OR1A1 and OR5AN1, that showed 10- to 100-fold more sensitivity to ligands as compared to previous assays. A single mouse can produce enough olfactory cilia for up to 4000 384-well assay wells, and isolated cilia can be stored frozen and thus preserved. This pipeline offers a sensitive and highly scalable ex vivo odor-screening platform that has the potential to decode human olfaction.

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Section: Discussionmentioning

confidence: 99%

A genetic platform for functionally profiling odorant receptors in olfactory cilia ex vivo

et al. 2022

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“…S5A). We recently characterized 5×21-OR1A1-Cherry (42) that reveals the dorsal epithelium is now expressing OR1A1 in 50% of the OSNs. Further increasing expression in the dorsal epithelium to 100% of the OSNs may double the signal.…”

Section: Discussionmentioning

confidence: 99%

A Genetic Platform for Functionally Profiling Odorant Receptors Ex Vivo Using Olfactory Cilia

Omura

Takabatake

Benjamin-Hong³

et al. 2021

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The molecular basis for odor perception in humans remains a black box as odorant receptors (ORs) are notoriously difficult to study outside of their native environment. Efforts toward OR expression and functional profiling in heterologous systems have been met with limited success due to poor efficiency of cell surface expression and consequently reduced G-protein signal amplification. We previously reported a genetic strategy in mice to increase the number of sensory neurons expressing specific ORs, which transforms the 10 million neurons of the mouse nose into a bioreactor producing large quantities of fully functional OR protein. We now describe the isolation of cilia from these bioreactors for two human ORs. Cilia are known to contain all components of the olfactory signal transduction machinery and can be placed into an ex vivo well-plate assay to rapidly measure robust, reproducible odor-specific responses. Our OR1A1 and OR5AN1 isolated cilia reveal 10-100fold more sensitivity than existing assays. Tissue from a single animal produces up to 4,000 384-well assay wells, and isolated olfactory cilia can be stored frozen and thus preserved for long term usage. This pipeline offers a sensitive, highly scalable ex vivo odor screening platform that opens the door for decoding human olfaction.

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Odorant Receptor Choice Mechanism Revealed by Analysis of a Highly Represented Odorant Receptor Transgene

Cited by 2 publications

References 32 publications

A genetic platform for functionally profiling odorant receptors in olfactory cilia ex vivo

A genetic platform for functionally profiling odorant receptors in olfactory cilia ex vivo

A Genetic Platform for Functionally Profiling Odorant Receptors Ex Vivo Using Olfactory Cilia

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