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B6;129S-Gt(ROSA)26Sortm39(CAG-hop/EYFP)Hze/J
Cat. No.: CEMM-07250291
Common Name: Ai39; Ai39(RCL-eNpHR3.0/EYFP)
These Ai39 mice conditionally express an improved Halorhodopsin/EYFP fusion protein from the endogenous Gt(ROSA)26Sor locus. Following Cre-mediated removal of the STOP cassette, EYFP expression is observed in the cre-expressing cells. Subsequent illumination of these cells with yellow-to-red light leads to reversible photoinhibition of action potential firing/neural activity in these cells. These Ai39 mice are useful for optogenetic studies to express an inhibitory opsin that effectively silences the activity of cortical neurons.
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| Status | Live Mouse Frozen Embryo |
| Age | 4 weeks 12 weeks Customized Age |
| Sex | Male Female |
| GENETICS | |
|---|---|
| Allele Symbol |
Gt(ROSA)26Sortm39(CAG-hop/EYFP)Hze
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| Allele Name |
targeted mutation 39
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| Allele Attributes |
Reporter
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| Gene Symbol |
Gt(ROSA)26Sor
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| Gene Name |
gene trap ROSA 26
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| Chromosome |
6
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| Expressed Genes |
HOP, Halorhodopsin, Natronomonas; YFP, Yellow Fluorescent Protein, jellyfish
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| MGI Accession ID | show more close |
| Site of Expression |
Cre excision of the stop signal results in expression of a halorhodopsin/EYFP (eNpHR3.0-EYFP) fusion protein in Cre-expressing tissues.
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| Strain of Origin |
(129S6/SvEvTac x C57BL/6NCrl)F1
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| Molecular Note |
The Rosa-CAG-LSL-eNpHR3.0-EYFP-WPRE targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), an FRT site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), an eNpHR3.0-EYFP fusion gene, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a BGH polyA signal, and an attB/attP-flanked PGK-FRT Neo-polyA cassette. The NpHR-EYFP fusion protein was first designed with the halorhodopsin from the halophilic bacterium Natronomas pharaonis (NpHR) fused in-frame with an enhanced yellow fluorescent protein (EYFP). To Create the eNpHR3.0-EYFP fusion protein, the NpHR-EYFP fusion protein was modified via addition of a membrane trafficking signal from the potassium channel Kir2.1 gene (amino acids KSRITSEGEYIPLDQIDINV) between NpHR and EYFP, as well as addition of an endoplasmic reticulum (ER) exporting sequence from the potassium channel Kir2.1 gene (amino acids FCYENEV) at the C-terminal of the EYFP. These modifications result in optimized expression in mammalian systems by preventing ER aggregation/enhancing membrane translocation, reducing bleb formation, and enhancing inhibitory capacity. The entire Rosa-CAG-LSL-eNpHR3.0-EYFP-WPRE targeting vector was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus.
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| HUSBANDRY | |
|---|---|
| Suggested Controls |
C57BL/6J
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| Breeding Considerations |
Heterozygous and homozygous mice are viable and fertile. When maintaining a live colony, heterozygous mice may be bred together, to wildtype mice from the colony or to C57BL/6J inbred mice. Alternatively, homozygous mice may be bred together.
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| Breeding Strategy |
Homozygote x Homozygote
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For Research Use Only.
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