These M72-IRES-ChR2-YFP knock-in mice express a ChR2(H134R)-EYFP fusion gene from the olfactory receptor 160 Olfr160 promoter. As such, the ChR2(H134R)-EYFP fusion protein is expressed olfactory sensory neurons (OSN) expressing this specific odorant receptor, M72. This targeted mutation strain may be useful for studying olfactory transduction, olfactory circuitry, and olfactory perception by light stimulation.
Thomas Bozza, Northwestern University
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter) | Olfr160 | olfactory receptor 160 |
These M72-IRES-ChR2-YFP mice express a ChR2(H134R)-EYFP fusion gene from the from the olfactory receptor 160 locus (Olfr160). Mice that are homozygous for the targeted mutation are viable and fertile. Only olfactory sensory neurons (OSN) expressing this specific odorant receptor, M72, express ChR2(H134R)-EYFP, rendering them light sensitive. Sensory neurons that express the modified allele express both ChR2-YFP as well as the intact M72 receptor and project axons to a medial and a lateral glomerulus on the dorsal surface of each olfactory bulb. Light directed at the glomerulus can elicit neuronal activity and drive odor guided behavior. These mutant mice thus allow precise, timed delivery of sensory input to the olfactory system. Homozygotes are viable and fertile. M72-IRES-ChR2-YFP mice are similar to STOCK Omptm1.1(COP4*/EYFP)Tboz/J mice (Stock No. 014173), which express ChR2(H134R)-EYFP all mature olfactory sensory neurons.
The ChR2(H134R)-EYFP fusion protein is composed of a Chlamydomonas reinhardtii-derived channelrhodopsin-2 that harbors a gain-of-function H134R substitution fused in-frame with an enhanced yellow fluorescent protein. The ChR2(H134R) is designed to cause larger stationary photocurrents compared to ChR2.
The bacterial opsins are retinal-binding proteins that combine a light-sensitive domain with an ion channel or pump; providing light-dependent ion transport, membrane potential alteration, and sensory functions to bacteria. This ChR2(H134R) functions as a blue light-driven cation channel that depolarizes the cell and causes action potentials. As such, illuminating ChR2(H134R)-expressing cells with blue light (450-490 nm) leads to rapid and reversible photostimulation of action potential firing activity in these cells.
A targeting vector was designed to insert an internal ribosome entry site IRES), the ChR2(H134R)-EYFP fusion gene, and a self-excising neo cassette, ACNF, downstream of the olfactory receptor 160 gene (Olfr160). This construct was electroporated into 129-derived embryonic stem (ES) cells and correctly targeted ES cells were aggregated with C57BL/6J morulae. Resulting blastocysts were implanted in pseudopregnant females. Chimeric male offspring were crossed to B6(Cg)-Tyrc-2J/J females (Stock No. 000058) to generate F1 heterozygous mice. These mice were bred together to generate homozygotes. Upon arrival at The Jackson Laboratory Repository, the mice were crossed to C57BL/6J mice (Stock No. 000664) for at least one generation to establish the colony. The Tyrc-2J allele was bred out of the colony.
Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish |
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Site of Expression | The ChR2(H134R)-EYFP fusion protein is expressed only in olfactory sensory neurons that express the specific odorant receptor, M72. |
Allele Name | targeted mutation 1.1, Thomas Bozza |
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Allele Type | Targeted (Reporter) |
Allele Synonym(s) | M72-ChR2; M72-ChR2-YFP; M72-IRES-ChR2-YFP |
Gene Symbol and Name | Olfr160, olfactory receptor 160 |
Gene Synonym(s) | |
Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish |
Site of Expression | The ChR2(H134R)-EYFP fusion protein is expressed only in olfactory sensory neurons that express the specific odorant receptor, M72. |
Strain of Origin | 129 |
Chromosome | 9 |
Molecular Note | A targeting vector was designed to insert an internal ribosome entry site (IRES), the ChR2(H134R)-EYFP fusion gene, and a self-excising neo cassette, ACNF, downstream of the olfactory receptor 160 gene. The neo cassette is excised. Only olfactory sensory neurons (OSN) expressing this specific odorant receptor, M72, express ChR2(H134R)-EYFP, rendering them light sensitive. |
When maintaining a live colony, homozygous mice may be bred together.
When using the M72-IRES-ChR2-YFP mouse strain in a publication, please cite the originating article(s) and include JAX stock #021206 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Olfr160<tm1.1(COP4*/EYFP)Tboz> |
Frozen Mouse Embryo | STOCK Olfr160<tm1.1(COP4*/EYFP)Tboz>/J | $2595.00 |
Frozen Mouse Embryo | STOCK Olfr160<tm1.1(COP4*/EYFP)Tboz>/J | $2595.00 |
Frozen Mouse Embryo | STOCK Olfr160<tm1.1(COP4*/EYFP)Tboz>/J | $3373.50 |
Frozen Mouse Embryo | STOCK Olfr160<tm1.1(COP4*/EYFP)Tboz>/J | $3373.50 |
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