This inducible calcium sensor transgene contains a conditional cardiac expressing eGFP-calmodulin fusion protein, GCaMP2. Expression of ccGCaMP2 is regulated by a tetracycline operator (tetO), driven by an enhanced Myh6 promoter which limits GCaMP2 expression in heart. These mice may be useful for measurement of myocyte Ca2+ transients in the beating heart of adult and embryonic mice in vivo and in vitro.
Michael I Kotlikoff, Cornell University
Genetic Background | Generation |
---|---|
|
Allele Type |
---|
Transgenic (Inducible, Inserted expressed sequence) |
This inducible calcium sensor transgene contains a conditional cardiac (cc) expressing eGFP-calmodulin fusion protein, GCaMP2. Mice homozygous for the ccGCaMP2 transgene are viable and fertile, normal in size and do not display any gross physical or behavioral abnormalities. Expression of ccGCaMP2 is regulated by a tetracycline operator (tetO), driven by an enhanced Myh6 promoter which limits GCaMP2 expression in heart. When bred with transgenic mice expressing a tetracycline transactivator (tTA) or reverse transactivator protein, cardiac-specific expression of GCaMP2 can be controlled by withdrawal or administration of tetracycline or a tetracycline analog, doxycycline, in the double mutant offspring. These mice may be useful for measurement of myocyte Ca2+ transients in the beating heart of adult and embryonic mice in vivo and in vitro.
This transgene contains a calcium-sensing molecule, GCaMP2, under the transcriptional control of a minimally active mouse alpha myosin heavy chain promoter (Myh6) fused to seven copies of the tetracycline-responsive promoter element (tetO). GCaMP2 consists of, from 5' to 3', a 13-residue peptide of myosin light chain kinase (M13), a circularly permutated eGFP interrupted at residue 145, calmodulin (CaM), and a polyA recognition sequence. Mutations resulting in increased brightness (D180Y and V93I), thermal stability (V163A, and S175G) and GFP dimerization prevention (A206K) were introduced. Three GATA sites and two thyroid-like response elements are ablated in Myh6 to maintain low levels of cardiac-specific expression and decreased background. This conditional cardiac GCaMP2 (ccGCaMP2) construct was injected into fertilized B6D2F2 oocytes and founder mice were bred to FVB/NTac mice to establish a colony. Transgenic offspring were also bred to C57BL/6J prior to arrival at The Jackson Laboratory. Upon arrival mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
Expressed Gene | GFP, Green Fluorescent Protein, |
---|---|
Expressed Gene | Mylk, myosin, light polypeptide kinase, mouse, laboratory |
Expressed Gene | Calm1, calmodulin 1, mouse, laboratory |
Site of Expression | When bred to mice expressing a tetracycline transactivator (tTA) or reverse transactivator (rtTA) these mice express GFP-calmodulin fusion protein, GCaMP2, in the heart when induced with doxycycline. |
Allele Name | transgene insertion 1, Michael I Kotlikoff |
---|---|
Allele Type | Transgenic (Inducible, Inserted expressed sequence) |
Allele Synonym(s) | ccGCaMP2 |
Gene Symbol and Name | Tg(Myh6*/tetO-GCaMP2)1Mik, transgene insertion 1, Michael I Kotlikoff |
Gene Synonym(s) | |
Promoter | Myh6, myosin, heavy polypeptide 6, cardiac muscle, alpha, murine, murine |
Promoter | tetO, tet operator, |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | Mylk, myosin, light polypeptide kinase, mouse, laboratory |
Expressed Gene | Calm1, calmodulin 1, mouse, laboratory |
Site of Expression | When bred to mice expressing a tetracycline transactivator (tTA) or reverse transactivator (rtTA) these mice express GFP-calmodulin fusion protein, GCaMP2, in the heart when induced with doxycycline. |
Strain of Origin | (C57BL/6 x DBA/2)F2 |
Chromosome | UN |
Molecular Note | This transgene contains a calcium-sensing molecule, GCaMP2, under the transcriptional control of a minimally active mouse alpha myosin heavy chain promoter fused to seven copies of the tetracycline-responsive promoter element (tetO). GCaMP2 consists of circularly permutated eGFP with the molecule interrupted at residue 145 and a 13-residue peptide of myosin light chain kinase (M13) and calmodulin (CaM), placed at the new N and C termini, respectively. An RSET polyHis peptide was introduced N-terminal to the original methionine in GCaMP1. Mutations resulting in increased brightness (D180Y and V93I), thermal stability (V163A, and S175G) and to prevent GFP dimerization (A206K) were introduced. GCaMP2 is 200 times brighter than GCaMP1. Three GATA sites and two thyroid-like response elements are ablated in Myh6 to make the promoter minimally active. When bred with transgenic mice expressing a tetracycline transactivator or reverse transactivator protein, cardiac-specific expression of GCaMP2 can be controlled by withdrawal or administration of a tetracycline analog. |
Mutations Made By | Michael Kotlikoff, Cornell University |
When maintaining a live colony, C57BL/6J females may be bred with heterozygous or homozygous males. The donating investigator reports some problems using homozygous females for breeding.
When using the STOCK Tg(Myh6*/tetO-GCaMP2)1Mik/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #012477 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Hemizygous or Non carrier for Tg(Myh6*/tetO-GCaMP2)1Mik/ |
Frozen Mouse Embryo | STOCK Tg(Myh6*/tetO-GCaMP2)1Mik/J | $2595.00 |
Frozen Mouse Embryo | STOCK Tg(Myh6*/tetO-GCaMP2)1Mik/J | $2595.00 |
Frozen Mouse Embryo | STOCK Tg(Myh6*/tetO-GCaMP2)1Mik/J | $3373.50 |
Frozen Mouse Embryo | STOCK Tg(Myh6*/tetO-GCaMP2)1Mik/J | $3373.50 |
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.
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.
What information were you hoping to find through your search?
How easy was it to find what you were looking for?
We may wish to follow up with you. Enter your email if you are happy for us to connect and reachout to you with more questions.
Please Enter a Valid Email Address
Thank you for sharing your feedback! We are working on improving the JAX Mice search. Come back soon for exciting changes.