These OMP-ChR2-YFP (OCY-58) knock-in mice express a ChR2(H134R)-EYFP fusion gene from the olfactory marker protein locus (Omp). As such, the ChR2(H134R)-EYFP fusion protein is expressed in all olfactory receptor neurons. 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, Null/Knockout) | Omp | olfactory marker protein |
These OMP-ChR2-YFP (OCY-58) mice express a ChR2(H134R)-EYFP fusion gene from the olfactory marker protein locus (Omp). Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. ChR2(H134R)-EYFP is expressed in all mature olfactory sensory neurons, rendering them light sensitive. Light directed at the olfactory epithelium or the glomeruli can elicit activity in mitral/tufted (M/T) cells of the olfactory bulb and can drive odor guided behavior. These mutant mice thus allow precise, timed delivery of stimuli input to the olfactory system. The Donating Investigator reports that this this targeted mutation creates a null allele. Due to possible olfactory deficits, analysis should be performed in heterozygous mice. The Donating Investigator recommends keeping the strain on the albino B6 background to avoid possible interference by pigmented melanocytes.
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 containing the ChR2(H134R)-EYFP fusion gene and a self-excising PGK-neo cassette, ACNF, was used to disrupt the targeted gene by replacing the coding sequence for Omp with ChR2(H134R)-EYFP coding sequence. The construct was electroporated into 129P2/OlaHsd-derived E14 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts, and chimeric animals were crossed to C57BL/6 mice. The resulting OMP-ChR2-YFP (OCY-58) mice were bred together to generate homozygotes.
Upon arrival at The Jackson Laboratory Repository, the mice were crossed to albino C57BL/6J mice (B6(Cg)-Tyrc-2J/J ; Stock No. 000058) for at least one generation to establish the colony. The Tyrc-2J allele was bred out of the colony.
Expressed Gene | COP4, Channelrhodopsin, Chlamydomonas |
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Site of Expression | YFP is expressed in all mature olfactory sensory neurons. |
Allele Name | targeted mutation 1.1, Thomas Bozza |
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Allele Type | Targeted (Reporter, Null/Knockout) |
Allele Synonym(s) | Omptm1.1(COP4*H134R/EYFP)Tboz; OMP-ChR2-YFP (OCY58) |
Gene Symbol and Name | Omp, olfactory marker protein |
Gene Synonym(s) | |
Expressed Gene | COP4, Channelrhodopsin, Chlamydomonas |
Site of Expression | YFP is expressed in all mature olfactory sensory neurons. |
Strain of Origin | 129P2/OlaHsd |
Chromosome | 7 |
Molecular Note | A targeting vector containing the COP4/EYFP, a modified channel rhodopsin-2 gene from Chlamydomonas reinhardtii (ChR2(H134R), and enhanced yellow fluorescent protein fusion gene, and the a self-excising PGK-neo cassette, ACNF, was used to disrupt the targeted gene. The neo cassette removed itself. |
Mutations Made By | Thomas Bozza, Northwestern University |
When maintaining a live colony, heterozygous mice may be bred to wildtype mice from the colony or to albino C57BL/6J mice (B6(Cg)-Tyrc-2J/J ; Stock No. 000058). The Donating Investigator recommends using the albino C57BL/6 background to avoid possible interference by pigmented melanocytes.
When using the STOCK Omptm1.1(COP4*/EYFP)Tboz/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #014173 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Omp<tm1(COP4/EYFP)Tboz> |
Frozen Mouse Embryo | STOCK Omp<tm1.1(COP4*/EYFP)Tboz>/J | $2595.00 |
Frozen Mouse Embryo | STOCK Omp<tm1.1(COP4*/EYFP)Tboz>/J | $2595.00 |
Frozen Mouse Embryo | STOCK Omp<tm1.1(COP4*/EYFP)Tboz>/J | $3373.50 |
Frozen Mouse Embryo | STOCK Omp<tm1.1(COP4*/EYFP)Tboz>/J | $3373.50 |
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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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