| |||||||||||||||||||
| These Ai39 mice conditionally express an improved Halorhodopsin/EYFP fusion protein (eNpHR3.0-EYFP) from the endogenous Gt(ROSA)26Sor locus. Expression is enhanced by the presence of a CAG promoter. Following Cre-mediated removal of the STOP cassette, eNpHR3.0-EYFP expression is observed in the cre-expressing cells. Subsequent illumination of these cells with yellow-to-red light (~589-680 nm) 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 (and perhaps other excitable cell types such as muscle cells and immune cells). | |||||||||||||||||||
- Description
- Disease & phenotype
- Genes & alleles
- Genotyping
- Health & care
- References
- Pricing & purchasing
- Terms of use
Description
Strain Information
Former Names B6;129S-Gt(ROSA)26Sortm39(CAG-HOP/EYFP)Hze/J (Changed: 01-FEB-13 ) B6;129S-Gt(ROSA)26Sortm39.1(CAG-HOP/EYFP)Hze/J (Changed: 15-JUL-11 ) Type Mutant Stock; Targeted Mutation; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Mating System Homozygote x Homozygote (Female x Male) 02-APR-12 Species laboratory mouse Generation F?+N1F4 (21-DEC-12)
Generation DefinitionsDonating Investigator Hongkui Zeng, Allen Institute for Brain Science (AIBS) Description
Ai39 mice heterozygous for the Rosa-CAG-LSL-eNpHR3.0-EYFP-WPRE conditional allele are viable and fertile. A loxP-flanked STOP cassette prevents transcription of the downstream eNpHR3.0-EYFP fusion gene (see below for detailed description of eNpHR3.0-EYFP). Because the CAG promoter driven reporter construct was targeted for insertion into the Gt(ROSA)26Sor locus, eNpHR3.0-EYFP expression is determined by which tissue(s) express Cre recombinase. When bred to mice that express Cre recombinase, the resulting offspring will have the STOP cassette deleted in the cre-expressing tissues; resulting in expression of the eNpHR3.0-EYFP fusion protein. The donating investigator reports that Ai39 mice do not express eNpHR3.0-EYFP prior to introduction of Cre recombinase. Fusion protein expression following exposure to cre can be detected by EYFP fluorescence and mRNA (in situ hybridization) [and presumably by antibody staining (immunohistochemistry); although this was not tested by the donating investigator]. Following exposure to Cre recombinase, illuminating eNpHR3.0-expressing neurons with yellow-to-red light leads to reversible photoinhibition of action potential firing/neural activity in these cells. The donating investigator specifically reports that expression of the inhibitory opsin occurs at levels sufficient to effectively silence the activity of cortical neurons. Fusion protein expression in tissues other than brain has not yet been evaluated by the donating investigator (April 2011). The phenotype of homozygous mice has not been characterized by the donating investigator. Of note, the FRT sites flanking the mutation allow for additional targeted replacement of the reporter sequences through Flp-mediated recombination if so desired. Similarly, the attB/attP-flanked selection cassette may be removed by introduction of the site-specific bacteriophage PhiC31 integrase if so desired.The eNpHR3.0-EYFP fusion protein is composed of the Natronomonas pharaonis-derived halorhodopsin (NpHR) fused in-frame with an enhanced yellow fluorescent protein (EYFP). The eNpHR3.0-EYFP fusion protein has been modified to harbor both the membrane trafficking signal and the endoplasmic reticulum (ER) exporting sequence from the potassium channel Kir2.1 gene. These improvements result in optimized expression in mammalian cells by preventing ER aggregation/enhancing membrane translocation, reducing bleb formation, and enhancing inhibitory capacity. The eNpHR3.0-EYFP fusion protein allows robust photocurrents/action potential inhibition when exposed to light ranging from yellow (~589 nm) to deep red (~660 nm) and at the red/infrared border (~680 nm).
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. The third-generation halorhodopsin eNpHR3.0 is an expression-optimized, yellow-to-red light-driven (~580-680 nm), inward chloride ion pump that causes hyperpolarization and prevents action potentials. For example, illumination of NpHR-expressing neurons leads to reversible photoinhibition of action potential firing/neural activity in these cells.
For characterization information, see images at the Allen Institute for Brain Science website (Ai39 images).
Development
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 Natronomonas 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 via electroporation of (129S6/SvEvTac x C57BL/6)F1-derived G4 embryonic stem (ES) cells. Correctly targeted ES cells (clone Ai39) were selected. Chimeric mice were bred with C57BL/6 wildtype mice and/or PhiC31-expressing mice (C57BL/6J congenic background; see Stock No. 007743). Mutant mice were bred with C57BL/6J wildtype mice for approximately three more generations prior to sending to The Jackson Laboratory Repository. Upon arrival, sperm from Ai39 males was used to fertilize oocytes from C57BL/6J inbred mice (Stock No. 000664) in order to rederive the living colony at The Jackson Laboratory Repository.All of the mice originally sent to The Jackson Laboratory Repository, as well as all the first generation of rederived offspring genotype as completely co-segregating for the neo cassette and eNpHR portion of the Ai39 construct. As such, these mice harbor the Ai39 mutation (retaining the attB/attP-flanked PGK-FRT-Neo-polyA cassette) rather than the Ai39Δneo mutation.
Control Information
| Control | ||
|---|---|---|
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
Related Strains
Allen Institute for Brain Science cre reporter strains
View Allen Institute for Brain Science cre reporter strains (16 strains)
012332 B6;SJL-Tg(Thy1-hop/EYFP)2Gfng/J 012334 B6;SJL-Tg(Thy1-hop/EYFP)4Gfng/J 017906 STOCK Tg(tetO-hop/EGFP,-COP4/mCherry)6Kftnk/J
Additional Web Information
Fluorescent Proteins/lacZ Systems
Introduction to Cre-lox technology
Phenotype
Phenotype Information
assigned by genotype
Gt(ROSA)26Sortm39(CAG-hop/EYFP)Hze/Gt(ROSA)26Sor+
involves: 129S6/SvEvTac * C57BL/6NCr
- no phenotypic analysis
- *normal* no phenotypic analysis (MGI Ref ID J:172634)
This mouse can be used to support research in many areas including:
YFP relatedNeurobiology Research
Cre-lox System
loxP-flanked Sequences
loxP-flanked Sequences: Test/Reporter
Fluorescent protein expression in neural tissue
Optogenetic and Pharmacogenetic tools
Cre-dependent optogenetic tools
Halorhodopsin expressing strains
Research Tools
Cre-lox System
loxP-flanked Sequences
loxP-flanked Sequences: Test/Reporter
Developmental Biology Research
Cre-lox System
transplantation marker for embryonic and adult tissue
Diabetes and Obesity Research
loxP
FLP-FRT System
FRT-flanked Sequences
FRT-flanked Sequences: Test/Reporter
Fluorescent Proteins
Genetics Research
Mutagenesis and Transgenesis
Mutagenesis and Transgenesis: Cre-lox System
Tissue/Cell Markers
Tissue/Cell Markers: Cre-lox System
Tissue/Cell Markers: cell marker for bone marrow transplantation
Tissue/Cell Markers: transplantation marker for embryonic and adult tissue
PhiC31 System
attB/attP-flanked Sequences
Reproductive Biology Research
Cre-lox System
transplantation marker for embryonic and adult tissue
Research Tools
Fluorescent Proteins
Genes & Alleles
Gene & Allele Information provided by MGI
| Allele Symbol | Gt(ROSA)26Sortm39(CAG-hop/EYFP)Hze | ||
|---|---|---|---|
| Allele Name | targeted mutation 39, Hongkui Zeng | ||
| Allele Type | Targeted (Reporter) | ||
| Common Name(s) | Ai39; Rosa-CAG-LSL-eNpHR3.0-EYFP; eNpHR3.0-EYFP; | ||
| Mutation Made By | Hongkui Zeng, Allen Institute for Brain Science (AIBS) | ||
| Strain of Origin | (129S6/SvEvTac x C57BL/6NCr)F1 | ||
| 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. | ||
| Expressed Gene | HOP, Halorhodopsin, Natronomonas | ||
| Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish | ||
| Yellow Fluorescent Protein (YFP) is a derivative of Green Fluorescent Protein (GFP), a versatile reporter molecule which has found use in many biological applications. The original molecule has been modified in order to enhance fluorescence intensity and shift the wavelength emitted when excited. When YFP is utilized in a transgenic construct, tissue expressing sufficient amounts of YFP will fluoresce yellowish-green when exposed to a 513 nm light source. | |||
| 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. [MGI Ref ID J:172634] [MGI Ref ID J:182204] | ||
| Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano | ||
| Chromosome | 6 | ||
| Gene Common Name(s) | AV258896; Gtrgeo26; Gtrosa26; R26; ROSA26; beta geo; expressed sequence AV258896; gene trap ROSA 26; gene trap ROSA b-geo 26; | ||
Genotyping
Genotyping Information
Genotyping Protocols
CAG-HOP/EYFP,Separated MCA
CAG-HOP/EYFP, Separated PCR
Helpful Links
Genotyping resources and troubleshooting
References
References provided by MGI
Selected Reference(s)
Madisen L; Mao T; Koch H; Zhuo JM; Berenyi A; Fujisawa S; Hsu YW; Garcia AJ 3rd; Gu X; Zanella S; Kidney J; Gu H; Mao Y; Hooks BM; Boyden ES; Buzsaki G; Ramirez JM; Jones AR; Svoboda K; Han X; Turner EE; Zeng H. 2012. A toolbox of Cre-dependent optogenetic transgenic mice for light-induced activation and silencing. Nat Neurosci 15(5):793-802. [PubMed: 22446880] [MGI Ref ID J:182204]
Zeng H. 2011. Direct Database Submission 2011/06/21_Dl1gAf MGI Direct Data Submission :. [MGI Ref ID J:172634]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number FGB29
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, heterozygous mice may be bred together, to wildtype siblings or to C57BL/6J inbred mice (Stock No. 000664). Breeding homozygous mice with heterozygous mice has been successful in producing heterozygous and homozygous offspring (March 2012). Mating System Homozygote x Homozygote (Female x Male) 02-APR-12 Diet Information LabDiet® 5K52/5K67
Pricing and Purchasing
Pricing, Supply Level & Notes, Controls
| Pricing for USA, Canada and Mexico shipping destinations |
|
Live Mice
Price per mouse (US dollars $) Gender Genotypes Provided Individual Mouse $195.00 Female or Male Homozygous for Gt(ROSA)26Sortm39(CAG-hop/EYFP)Hze
Price per Pair (US dollars $) Pair Genotype $390.00 Homozygous for Gt(ROSA)26Sortm39(CAG-hop/EYFP)Hze x Homozygous for Gt(ROSA)26Sortm39(CAG-hop/EYFP)Hze Standard Supply
Repository-Live.
Repository-Live represents an exclusive set of over 1500 unique mouse models across a vast array of research areas. Breeding colonies provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. If a Repository strain is not immediately available, then within 2 to 3 business days, you will receive an estimated availability timeframe for your inquiry or order along with various delivery options. Repository strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping. We will note and try to accommodate requests for specific ages of Repository strains but cannot guarantee provision of these strains at specific ages. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, please let us know.
| Pricing for International shipping destinations |
|
Live Mice
Price per mouse (US dollars $) Gender Genotypes Provided Individual Mouse $253.50 Female or Male Homozygous for Gt(ROSA)26Sortm39(CAG-hop/EYFP)Hze
Price per Pair (US dollars $) Pair Genotype $507.00 Homozygous for Gt(ROSA)26Sortm39(CAG-hop/EYFP)Hze x Homozygous for Gt(ROSA)26Sortm39(CAG-hop/EYFP)Hze Standard Supply
Repository-Live.
Repository-Live represents an exclusive set of over 1500 unique mouse models across a vast array of research areas. Breeding colonies provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. If a Repository strain is not immediately available, then within 2 to 3 business days, you will receive an estimated availability timeframe for your inquiry or order along with various delivery options. Repository strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping. We will note and try to accommodate requests for specific ages of Repository strains but cannot guarantee provision of these strains at specific ages. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, please let us know.
|
|
|
Standard Supply
Repository-Live.
Repository-Live represents an exclusive set of over 1500 unique mouse models across a vast array of research areas. Breeding colonies provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. If a Repository strain is not immediately available, then within 2 to 3 business days, you will receive an estimated availability timeframe for your inquiry or order along with various delivery options. Repository strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping. We will note and try to accommodate requests for specific ages of Repository strains but cannot guarantee provision of these strains at specific ages. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, please let us know.
Control Information
| Control | ||
|---|---|---|
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
| Control Pricing Information for Genetically Engineered Mutant Strains. | ||
Payment Terms and Conditions
Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.
See Terms of Use tab for General Terms and Conditions
The Jackson Laboratory's Genotype Promise
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.Ordering Information
JAX® Mice
Surgical and Preconditioning Services
JAX® Services
Customer Services and Support
Tel: 1-800-422-6423 or 1-207-288-5845
Fax: 1-207-288-6150
Technical Support Email Form
Terms of Use
Terms of Use
General Terms and Conditions
For Licensing and Use Restrictions view the link(s) below:
- Strain(s) not available to companies or for-profit entities.
- Strain(s) not available to companies or for-profit entities.
Contact information
General inquiries regarding Terms of UseContracts Administration
| phone: | 207-288-6470 |
| fax: | 207-288-6655 |
JAX® Mice, Products & Services Conditions of Use
"MICE" means mouse strains, their progeny derived by inbreeding or crossbreeding, unmodified derivatives from mouse strains or their progeny supplied by The Jackson Laboratory ("JACKSON"). "PRODUCTS" means biological materials supplied by JACKSON, and their derivatives. "RECIPIENT" means each recipient of MICE, PRODUCTS, or services provided by JACKSON including each institution, its employees and other researchers under its control. MICE or PRODUCTS shall not be: (i) used for any purpose other than the internal research, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services. Acceptance of MICE or PRODUCTS from JACKSON shall be deemed as agreement by RECIPIENT to these conditions, and departure from these conditions requires JACKSON's prior written authorization.No Warranty
MICE, PRODUCTS AND SERVICES ARE PROVIDED “AS IS”. JACKSON EXTENDS NO WARRANTIES OF ANY KIND, EITHER EXPRESS, IMPLIED, OR STATUTORY, WITH RESPECT TO MICE, PRODUCTS OR SERVICES, INCLUDING ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, OR ANY WARRANTY OF NON-INFRINGEMENT OF ANY PATENT, TRADEMARK, OR OTHER INTELLECTUAL PROPERTY RIGHTS.
In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of mice, products or services, JACKSON will, at its option, provide credit or replacement for the mice or product received or the services provided.
No Liability
In no event shall JACKSON, its trustees, directors, officers, employees, and affiliates be liable for any causes of action or damages, including any direct, indirect, special, or consequential damages, arising out of the provision of MICE, PRODUCTS or services, including economic damage or injury to property and lost profits, and including any damage arising from acts or negligence on the part of JACKSON, its agents or employees. Unless prohibited by law, in purchasing or receiving MICE, PRODUCTS or services from JACKSON, purchaser or recipient, or any party claiming by or through them, expressly releases and discharges JACKSON from all such causes of action or damages, and further agrees to defend and indemnify JACKSON from any costs or damages arising out of any third party claims.
MICE and PRODUCTS are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.
The foregoing represents the General Terms and Conditions applicable to JACKSON’s MICE, PRODUCTS or services. In addition, special terms and conditions of sale of certain MICE, PRODUCTS or services may be set forth separately in JACKSON web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, PRODUCTS and services by JACKSON, and by its licensees and distributors.
Acceptance of delivery of MICE, PRODUCTS or services shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on JACKSON, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, PRODUCTS or services by JACKSON.