Strain Name:

STOCK Tg(tetO-CHRM3*)1Blr/J

Stock Number:

014093

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Availability:

Repository- Live

Use Restrictions Apply, see Terms of Use
These TRE-hM3Dq transgenic mice may be bred to generate Tet-Off/Tet-On mutant mice with conditional (inducible/reversible) expression of hM3Dq; a mutant G protein-coupled receptor (GPCR) that activates the canonical Gq pathway specifically following administration of the pharmacologically inert molecule clozapine-N-oxide (CNO). TRE-hM3Dq transgenic mice may be useful to study receptor-specific functions or general downstream cellular signaling emanating from the activated GPCR.

Description

Strain Information

Type Mutant Stock; Transgenic;
Additional information on Genetically Engineered and Mutant Mice.
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Mating SystemHemizygote x Noncarrier         (Female x Male)   16-DEC-11
Specieslaboratory mouse
GenerationF?+F3 (08-JAN-13)
Generation Definitions
 
Donating Investigator Bryan L Roth,   University of North Carolina at Chapel Hill

Description
Hemizygous TRE-hM3Dq transgenic mice are viable and fertile, with no reported phenotypic abnormalities. The TRE-hM3Dq transgene has a modified Tet response element (TRE or tetO) upstream of the mutant G protein-coupled receptor, hM3Dq (a human muscarinic 3 receptor with two amino acid substitutions (Y149C3.33/A239G5.46) that abolish receptor affinity for the native ligand, acetylcholine (ACh), but allow receptor binding and subsequent activation by the small pharmacologically inert molecule clozapine-N-oxide (CNO)). When bred with another mouse expressing tetracycline-controlled transactivator protein (tTA) or reverse tetracycline-controlled transactivator protein (rtTA), hM3Dq expression in the resulting double mutant offspring can be regulated with tetracycline or its analog doxycycline (dox). As designed, TRE-hM3Dq transgenic mice have no reported levels of hM3Dq activity in tTA(+dox) or rtTA(-dox) cell types. In TRE-hM3Dq transgenic tTA(-dox) or TRE-hM3Dq transgenic rtTA(+dox) cells types, hM3Dq expression results in activation of the canonical Gq pathway only following administration of CNO.

These TRE-hM3Dq transgenic mice may be useful to study receptor-specific functions or general downstream cellular signaling emanating from the activated G protein-coupled receptor (GPCR). For example, when bred to a strain expressing tTA in forebrain neurons (Camk2a-tTA; see Stock No. 003010), these transgenic mice are a model allowing in vivo chemical control of neuronal activity, neuronal firing, and non-neuronal signaling.

In addition, breeding TRE-hM3Dq mice with cfos-htTA mice (Stock No. 018306) generates double transgenic offspring with hM3Dq expression in excitatory neurons that are sufficiently active to drive the c-fos promoter. Those hM3Dqfos double transgenic mice allow CNO ligand-induced, artificial reactivation of neurons associated with a specific memory. Temporal modulation may also be employed using dox. Such hM3Dqfos double transgenic mice allow one to generate and artificially-activate a synthetic memory trace, and may be useful in studying the spatial pattern of activity at the time of learning and retrieval.

Of note, several pharmacogenetic tool strains from the donating investigator are available from The Jackson Laboratory Repository; including these Tet-responsive TRE-hM3Dq transgenic mice (Stock No. 014093), the adora2A-rM3Ds transgenic mice (Stock No. 017863) and the Tet-responsive TRE-hM4Di transgenic mice (Stock No. 024114).

Development
The TRE-HA-hM3Dq transgene was designed with a modified Tet response element (TRE or (tetO); described in detail below), a hemagglutinin epitope tag (HA), the hM3Dq sequence, and an SV40 polyA signal. The hM3Dq sequence is a Gq-coupled human M3 muscarinic DREADD (designer receptor exclusively activated by designer drug). To create the hM3Dq sequence, the wildtype human muscarinic 3 receptor (CHRM3) sequence was modified via site-directed mutagenesis to harbor two amino acid substitutions (Y149C3.33/A239G5.46) that abolish receptor affinity for the native ligand, acetylcholine (ACh), but allow receptor binding and subsequent activation by the small drug-like molecule clozapine-N-oxide (CNO).
This 2.36 kb TRE-HA-hM3Dq transgene was injected into the pronuclei of B6SJL hybrid mouse oocytes. Founder TRE-hM3Dq mice (line 1) were bred with B6;CBA-Tg(Camk2a-tTA)1Mmay/J (see Stock No. 003010) to establish the colony. TRE-hM3Dq transgenic mice were bred together or to wildtype (noncarrier) mice from the colony for approximately eight generations, and then crossed with C57BL/6J mice for one or two generations. The Camk2a-tTA transgene was removed during these breedings. Upon arrival at The Jackson Laboratory Repository, TRE-hM3Dq transgenic mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Additional information on hM3Dq:
More information on hM3Dq or other DREADDs may be available at the University of North Carolina Designer Receptors Exclusively Activated by Designer Drugs DREADD wiki webpage.

Additional information on TRE promoter:
The TRE promoter used in these transgenic mice is the Tet-responsive Ptight promoter. This Ptight promoter contains a modified Tet response element (TREmod), which consists of seven direct repeats of a 36-bp sequence each containing the 19-bp tet operator sequence (tetO). The TREmod is just upstream of a minimal human cytomegalovirus (CMV) promoter (PminCMVΔ), which lacks the enhancer that is part of the complete CMV promoter. Consequently, Ptight is silent in the absence of binding of TetR or rTetR to the tetO sequences.

Control Information

  Control
   Noncarrier
   100011 B6CBAF1/J (approximate)
 
  Considerations for Choosing Controls

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017330   B6.Cg-Tg(tetO-TAg*)175Kndl/J
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016571   FVB-Tg(Myh6/tetO-Gata6)2Jmol/J
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012684   FVB-Tg(Myh6/tetO-POSTN)22.1Jmol/J
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013156   FVB-Tg(tetO-CDK5R1*)1Vln/J
013777   FVB-Tg(tetO-Cacna1g)1Jmol/J
013778   FVB-Tg(tetO-Cacnb2)1Jmol/J
013779   FVB-Tg(tetO-Cacnb2)2Jmol/J
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010578   FVB-Tg(tetO-Dusp6)1Jmol/J
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012387   FVB-Tg(tetO-Ppargc1a)1Dpk/J
012385   FVB-Tg(tetO-Ppargc1b)7Dpk/J
022979   FVB-Tg(tetO-Thbs4)17.7Jmol/J
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019038   FVB.Cg-Tg(tetO-GLI1)10Rup/Mmjax
019039   FVB.Cg-Tg(tetO-KLF4)32831Rup/Mmjax
008244   FVB.Cg-Tg(tetO-cre)1Jaw/J
012459   FVB/N-Tg(Myh6*/tetO-Capn1)L2Gwd/J
005941   FVB/N-Tg(tetO-Aurkb,lacZ)41Kra/J
006202   FVB/N-Tg(tetO-BCR/ABL1)2Dgt/J
014547   FVB/N-Tg(tetO-Fasl)BDepa/J
019376   FVB/N-Tg(tetO-MYC)36aBop/J
022938   FVB/N-Tg(tetO-Wnt5a)17Rva/J
003315   FVB/N-Tg(tetORo1-lacZ)3Conk/J
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006999   STOCK Dbttm1Geh Tg(Cebpb-tTA)5Bjd Tg(tetO-DBT)A1Geh/J
011004   STOCK Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm3(tetO-Pou5f1,-Sox2,-Klf4,-Myc)Jae/J
011011   STOCK Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm4(tetO-Pou5f1,-Sox2,-Klf4,-Myc)Jae/J
011013   STOCK Gt(ROSA)26Sortm1(rtTA*M2)Jae Col1a1tm5(tetO-Pou5f1,-Klf4,-Myc)Jae/J
018999   STOCK Gt(ROSA)26Sortm1(tTA,tetO-Mir155)Fjsl/J
017596   STOCK Gt(ROSA)26Sortm1.1(rtTA,EGFP)Nagy Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN2*delta7)4299Ahmb Tg(tetO-SMN2,-luc)#aAhmb/J
017597   STOCK Gt(ROSA)26Sortm1.1(rtTA,EGFP)Nagy Tg(SMN2)89Ahmb Smn1tm1Msd Tg(SMN2*delta7)4299Ahmb Tg(tetO-SMN2,-luc)#bAhmb/J
024854   STOCK Tg(Camk2a-tTA)1Mmay Tg(tetO-MAPT*P301L)#Kha/J
008755   STOCK Tg(Ins2-rtTA)2Efr Tg(teto-DTA)1Gfi/J
012477   STOCK Tg(Myh6*/tetO-GCaMP2)1Mik/J
016572   STOCK Tg(Myh6/tetO-Gata4)1Jmol/J
014544   STOCK Tg(tetO-ABL1*P242E*P249E)CPdav/J
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008168   STOCK Tg(tetO-DTA)1Gfi/J
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024509   STOCK Tg(tetO-Gata6)1Abl/J
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005699   STOCK Tg(tetO-Ipf1,EGFP)956.6Macd/J
005728   STOCK Tg(tetO-Ipf1,lacZ)958.1Macd/J
012441   STOCK Tg(tetO-LRRK2*G2019S)E3Cai/J
017918   STOCK Tg(tetO-MAML1*/EGFP)2Akar/J
017599   STOCK Tg(tetO-SMN2,-luc)#aAhmb/J
017600   STOCK Tg(tetO-SMN2,-luc)#bAhmb/J
012442   STOCK Tg(tetO-SNCA*A53T)E2Cai/J
006224   STOCK Tg(tetO-cre)1Jaw/J
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012449   STOCK Tg(teto-LRRK2)C7874Cai/J
View Strains carrying other alleles of tetO     (133 strains)

Additional Web Information

Tet Expression Systems

Phenotype

Phenotype Information

View Related Disease (OMIM) Terms

Related Disease (OMIM) Terms provided by MGI
- Potential model based on transgenic expression of an ortholog of a human gene that is associated with this disease. Phenotypic similarity to the human disease has not been tested.
Abdominal Muscles, Absence of, with Urinary Tract Abnormality and Cryptorchidism   (CHRM3)
View Research Applications

Research Applications
This mouse can be used to support research in many areas including:

Cancer Research
Growth Factors/Receptors/Cytokines

Cell Biology Research
Signal Transduction

Diabetes and Obesity Research
Insulin Receptors and Growth Factors

Immunology, Inflammation and Autoimmunity Research
Growth Factors/Receptors/Cytokines
Intracellular Signaling Molecules

Neurobiology Research
Optogenetic and Pharmacogenetic tools
      Pharmacogenetic tools
Receptor Defects
      G protein-coupled receptor
Tet Expression System
      tTA/rtTA Responsive Strains

Research Tools
Cancer Research
      Tetop Tet System
Cardiovascular Research
      Tetop Tet System
Genetics Research
      Mutagenesis and Transgenesis: Tetop Tet System
Neurobiology Research
      Tetop Tet System
Reproductive Biology Research
      Tetop Tet System
Tet Expression Systems
      tTA/rtTA Responsive Strains

Genes & Alleles

Gene & Allele Information provided by MGI

 
Allele Symbol Tg(tetO-CHRM3*)1Blr
Allele Name transgene insertion 1, Bryan Roth
Allele Type Transgenic (Inducible, Inserted expressed sequence)
Common Name(s) TRE-HA-hM3Dq; TRE-hM3Dq; tetO-hM3Dq;
Mutation Made By Bryan Roth,   University of North Carolina at Chapel Hill
Strain of Origin(B6 x SJL)F1
Expressed Gene CHRM3, cholinergic receptor, muscarinic 3, human
Promoter tetO, tet operator,
Promoter CMV, cytomegalovirus, human
Molecular Note The Tet-responsive P promoter which consists of seven direct repeats of 36-bp sequence each containing the 19-bp tet operator sequence (tetO) is just upstream of a minimal human cytomegalovirus (CMV) promoter. This is upstream of a hemagglutinin epitoptag and a Gq-coupled human M3 muscarinic receptor. The wildtype human muscarinic 3 receptor sequence was modified via site-directed mutagenesis to harbor two amino acid substitutions (Y149C and A239G) that abolish receptor affinity for the native ligand,acetylcholine (ACh), but allow receptor binding and subsequent activation by the small drug-like molecule clozapine-N-oxide (CNO). [MGI Ref ID J:154952]
 
 
 

Genotyping

Genotyping Information

Genotyping Protocols

** human CHRM3*A239G, Pyrosequencing
** human CHRM3*Y149C, Pyrosequencing
Tg(tetO-CHRM3*)1Blr, High Resolution Melting
Tg(tetO-CHRM3*)1Blr, Standard PCR
Tg(tetO-CHRM3*)1Blr, Separated PCR


Helpful Links

Genotyping resources and troubleshooting

References

References provided by MGI

Selected Reference(s)

Alexander GM; Rogan SC; Abbas AI; Armbruster BN; Pei Y; Allen JA; Nonneman RJ; Hartmann J; Moy SS; Nicolelis MA; McNamara JO; Roth BL. 2009. Remote control of neuronal activity in transgenic mice expressing evolved G protein-coupled receptors. Neuron 63(1):27-39. [PubMed: 19607790]  [MGI Ref ID J:154952]

Additional References

Tg(tetO-CHRM3*)1Blr related

Garner AR; Rowland DC; Hwang SY; Baumgaertel K; Roth BL; Kentros C; Mayford M. 2012. Generation of a synthetic memory trace. Science 335(6075):1513-6. [PubMed: 22442487]  [MGI Ref ID J:181832]

Health & husbandry

Health & Colony Maintenance Information

Animal Health Reports

Room Number           AX18

Colony Maintenance

Breeding & HusbandryWhen maintaining a live colony, transgene carrier mice may be bred together or to wildtype (noncarrier) mice from the colony. The donating investigator reports they have not tried to generate homozygous mice (November 2011).
Mating SystemHemizygote x Noncarrier         (Female x Male)   16-DEC-11
Diet Information LabDiet® 5K52/5K67

Pricing and Purchasing

Pricing, Supply Level & Notes, Controls


Pricing for USA, Canada and Mexico shipping destinations View International Pricing

Live Mice

Price per mouse (US dollars $)GenderGenotypes Provided
Individual Mouse $232.00Female or MaleHemizygous for Tg(tetO-CHRM3*)1Blr  
Price per Pair (US dollars $)Pair Genotype
$304.00Hemizygous for Tg(tetO-CHRM3*)1Blr x Noncarrier  
$304.00Noncarrier x Hemizygous for Tg(tetO-CHRM3*)1Blr  

Standard Supply

Repository-Live.
Repository-Live represents an exclusive set of over 1800 unique mouse models across a vast array of research areas. Breeding colonies provide mice for large and small orders and fluctuate in size depending on current research demand. If a strain is not immediately available, you will receive an estimated availability timeframe for your inquiry or order in 2-3 business days. Repository strains typically are delivered at 4 to 8 weeks of age. Requests for specific ages will be noted but not guaranteed and we do not accept age requests for breeder pairs. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, we will do our best to accommodate your age request.

Pricing for International shipping destinations View USA Canada and Mexico Pricing

Live Mice

Price per mouse (US dollars $)GenderGenotypes Provided
Individual Mouse $301.60Female or MaleHemizygous for Tg(tetO-CHRM3*)1Blr  
Price per Pair (US dollars $)Pair Genotype
$395.20Hemizygous for Tg(tetO-CHRM3*)1Blr x Noncarrier  
$395.20Noncarrier x Hemizygous for Tg(tetO-CHRM3*)1Blr  

Standard Supply

Repository-Live.
Repository-Live represents an exclusive set of over 1800 unique mouse models across a vast array of research areas. Breeding colonies provide mice for large and small orders and fluctuate in size depending on current research demand. If a strain is not immediately available, you will receive an estimated availability timeframe for your inquiry or order in 2-3 business days. Repository strains typically are delivered at 4 to 8 weeks of age. Requests for specific ages will be noted but not guaranteed and we do not accept age requests for breeder pairs. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, we will do our best to accommodate your age request.

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Standard Supply

Repository-Live.
Repository-Live represents an exclusive set of over 1800 unique mouse models across a vast array of research areas. Breeding colonies provide mice for large and small orders and fluctuate in size depending on current research demand. If a strain is not immediately available, you will receive an estimated availability timeframe for your inquiry or order in 2-3 business days. Repository strains typically are delivered at 4 to 8 weeks of age. Requests for specific ages will be noted but not guaranteed and we do not accept age requests for breeder pairs. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, we will do our best to accommodate your age request.

Control Information

  Control
   Noncarrier
   100011 B6CBAF1/J (approximate)
 
  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.
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Tel: 1-800-422-6423 or 1-207-288-5845
Fax: 1-207-288-6150
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Terms of Use

Terms of Use


General Terms and Conditions


For Licensing and Use Restrictions view the link(s) below:
- Use of MICE by companies or for-profit entities requires a license prior to shipping.

Contact information

General inquiries regarding Terms of Use

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phone:207-288-6470

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.


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