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B6.Cg-Gt(ROSA)26Sortm40.1(CAG-aop3/EGFP)Hze/J
Cat. No.: CEMM-07250437
Common Name: Ai40(RCL-ArchT/EGFP)-D; Ai40D
These Ai40D mice conditionally express an improved Archaerhodopsin-3/EGFP fusion protein (ArchT-EGFP) from the endogenous Gt(ROSA)26Sor locus. Expression is enhanced by the presence of a CAG promoter. Following Cre-mediated removal of the floxed-STOP cassette, ArchT-EGFP expression is observed in the cre-expressing cells. Subsequent illumination of these cells with yellow-green light (~575 nm) leads to reversible photoinhibition of action potential firing/neural activity in these cells. These Ai40D mice are useful for optogenetic studies to express an inhibitory opsin that effectively silences the activity of cortical neurons (and perhaps other excitable cell types such as muscle cells and immune cells). ArchT has greater than 3-fold improvement in light sensitivity compared to Arch, equating to greater than 200% increase in brain tissue volume addressed by a typical single optical fiber.
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| Status | Live Mouse Frozen Embryo |
| Age | 4 weeks 12 weeks Customized Age |
| Sex | Male Female |
| GENETICS | |
|---|---|
| Allele Symbol |
Gt(ROSA)26Sortm40.1(CAG-aop3/EGFP)Hze
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| Allele Name |
targeted mutation 40.1
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| Allele Attributes |
Conditional ready (e.g. floxed); 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 |
GFP, Green Fluorescent Protein; AOP3, Archaerhodopsin, Halorubrum sodomense
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| MGI Accession ID | show more close |
| Site of Expression |
These mice conditionally express an improved Archaerhodopsin-3/EGFP fusion protein (ArchT-EGFP). ArchT expresses extremely well on the membranes of neurons, including good expression on axons,
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| Strain of Origin |
(129S6/SvEvTac x C57BL/6NCrl)F1
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| Molecular Note |
The Rosa-pCAG-LSL-ArchT/EGFP-WPRE-bGHpA-AttB-PGK-FRT Neo-polyA-AttP targeting vector was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (frompCAGGS), an FRT site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames linked to three repeats of an SV40 polyA signal), an ArchT-EGFP fusion gene, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a bovine growth hormone polyA signal, an attB site, a PGK promoter, a second FRT site, neomycin resistance gene-SV40polyA signal, and an attP site. The ArchT-EGFP fusion gene is a mammalian codon-optimized cDNA sequence encoding the Archaerhodopsin-3 protein derived from the halophilic bacterium Halorubrum strain TP009 (ArchT or aR-TP009) fused in-frame via a 15 nucleotide linker to the amino terminus of an enhanced green fluorescent protein (EGFP). PhiC31-mediated recombination removed the attB/attP-flanked PGK-FRT Neo-polyA cassette and left a recombined attB/attP site (attL).
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| HUSBANDRY | |
|---|---|
| Suggested Controls |
C57BL/6J
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| Breeding Considerations |
When maintaining the live colony, heterozygous mice may be bred to wildtype mice from the colony or to C57BL/6J inbred mice. Alternatively, homozygous mice may be bred togather.
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| Breeding Strategy |
Homozygote x Homozygote
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For Research Use Only.
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