Overview

Also Known As:Cdh5-CatCh-IRES-lacZ , Cdh5^BAC-CatCh-IRES-lacZ

Cdh5-CatCh-IRES-lacZ transgenic mice express calcium-translocating channelrhodopsin CatCh (an L132C mutant channelrhodopsin with enhanced Ca2+ permeability, accelerated response kinetics and ~70-fold greater light sensitivity) in vascular endothelial cells (e.g., heart, lung, brain). Illumination of the CatCh molecule with blue-to-green light in vascular endothelial cells is expected to result in reversible photostimulation of channel opening, calcium inflow and muscle relaxation.

Donating Investigator

Michael I Kotlikoff, Cornell University

Genetic overview

Genetic Background	Generation

Tg(Cdh5-COP4*L132C,-lacZ)B28-2Mik

Allele Type
Transgenic (Reporter)

View Genetics

Research Applications

Research Tools
Neurobiology Research
Cardiovascular Research
Developmental Biology Research
Internal/Organ Research

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

Cdh5-CatCh-IRES-lacZ transgenic mice express the calcium-translocating channelrhodopsin CatCh and lacZ under control of the Cdh5 locus promoter/enhancer regions within the BAC transgene. Cdh5-CatCh-IRES-lacZ mice from founder line B28-2 exhibit high expression levels of CatCh and lacZ in vascular endothelial cells (e.g., heart, lung, brain) - consistent with endogenous Cdh5. Additional details described further below.

CatCh is a calcium-translocating channelrhodopsin variant designed to have enhanced Ca2+ permeability, accelerated response kinetics and ~70-fold greater sensitivity to blue-to-green light (~450-530 nm; max 474 nm). Additional CatCh information is described below. For Cdh5-CatCh-IRES-lacZ transgenic mice, illumination of the CatCh molecule with this light in vascular endothelial cells is expected to result in reversible photostimulation of channel opening, calcium inflow and relaxation of the muscle.

The donating investigator reports that expression patterns appear to mimic the endogenous Cdh5 expression. Specifically, lacZ staining was observed in endothelial lined capillaries and vessels of the heart, lung and brain. Also, high levels of lacZ expression were observed in diaphragm vasculature. CatCh activation with light has not specifically been tested for Cdh5-CatCh-IRES-lacZ transgenic mice to date (July 2019) - although CatCh function has been shown to work as expected for other CHROMus lines (e.g., Stock Nos. 030334, 033344, etc.).

Mice hemizygous for the Cdh5-CatCh-IRES-lacZ transgene are viable and fertile with normal breeding, and no reported gross phenotypic abnormalities. To date (July 2019), it has not been attempted to make this strain homozygous.

This mouse model is available by way of a collaborative effort between Cornell/National Heart Lung Blood Resource for Optogenetic Mouse Signaling (CHROMus) and The Jackson Laboratory.

Development

Cdh5-CatCh-IRES-lacZ transgenic mice (also called Cdh5^BAC-CatCh-IRES-lacZ) were designed in the laboratory of Dr. Michael I. Kotlikoff (Cornell University) as part of Cornell/National Heart Lung Blood Resource for Optogenetic Mouse Signaling (CHROMus).
The ~200 kbp C57BL/6J mouse bacterial artificial chromosome (BAC) RP23-453P1 was obtained; containing the entire cadherin 5 locus (Cdh5) as well as ~124 kbp of 5' flanking sequences (including the complete Gm32728, Gm39236 and Gm8748 predicted gene loci) and ~33 kbp of 3' flanking sequences (including a 5' portion of the Bean1 locus). Using homologous recombination/BAC recombineering, a 5640 bp CatCh-IRES-lacZ-pA construct (CatCh, internal ribosome entry site, β-galactosidase and an SV40 polyadenylation signal) was inserted into the ATG start site of the BAC Cdh5 gene (replacing the initiation codon of Cdh5 in exon 2). No other loci on the BAC were altered. The 11.6 kbp BAC vector (pBACe3.6) contained a chloramphenicol selection cassette.
The resulting ~217.5 kbp modified BAC (Cdh5^BAC-CatCh-IRES-lacZ) was purified and then microinjected into the male pronucleus of C57BL/6J x SJL/J embryos. Founder animals were bred to C57BL/6J inbred mice for germline transmission. Cdh5-CatCh-IRES-lacZ founder line B28-2 was identified with high effector and lacZ expression in vascular endothelial cells. The donating investigator reports that Cdh5-CatCh-IRES-lacZ transgenic mice from this founder line were backcrossed to C57BL/6J for a total of at least five generations, and then hemizygous males (black coat color) were sent to The Jackson Laboratory Repository in 2019. Upon arrival, sperm was cryopreserved. To establish our living colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J inbred females (Stock No. 000664).

Of note, the donating investigator reports that, at least once during backcrossing, a hemizygous female was bred to a C57BL/6J inbred male (thus the Y chromosome of the congenic strain is of C57BL/6J origin). The transgene insertion site(s) and transgene copy number were not characterized (July 2019).

The calcium translocating channelrhodopsin CatCh [ChR2(L132C)] cDNA sequence used here encodes the first 309 amino acids of green alga Chlamydomonas reinhardtii channelrhodopsin-2 (ChR2 or cop4) that has been modified with an L132C amino acid substitution (CTC-to-TGC) designed to cause enhanced Ca2+ permeability.

Control Suggestions

Noncarrier

Approximate Controls

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Genetics

Tg(Cdh5-COP4*L132C,-lacZ)B28-2Mik

Allele Symbol: Tg(Cdh5-COP4*L132C,-lacZ)B28-2Mik

Allele Name	transgene insertion B28-2, Michael I Kotlikoff
Allele Type	Transgenic (Reporter)
Allele Synonym(s)	Tg(Cdh5-COP4*L132C,-lacZ)B28-2Mik; transgene insertion B28-2, Michael I Kotlikoff
Gene Symbol and Name	Tg(Cdh5-COP4*L132C,-lacZ)B28-2Mik, transgene insertion B28-2, Michael I Kotlikoff
Gene Synonym(s)
Strain of Origin	C57BL/6J x SJL/J
Chromosome	UN
Molecular Note	Cdh5-CatCh-IRES-lacZ transgenic mice (also called Cdh5-CatCh-IRES-lacZ) were designed in the laboratory of Dr. Michael I. Kotlikoff (Cornell University) as part of Cornell/National Heart Lung Blood Resource for Optogenetic Mouse Signaling (CHROMus). The ~200 kbp C57BL/6J mouse bacterial artificial chromosome (BAC) RP23-453P1 was obtained; containing the entire cadherin 5 locus (Cdh5) as well as ~124 kbp of 5' flanking sequences (including the complete Gm32728, Gm39236 and Gm8748 predicted gene loci) and ~33 kbp of 3' flanking sequences (including a 5' portion of the Bean1 locus). Using homologous recombination/BAC recombineering, a 5640 bp CatCh-IRES-lacZ-pA construct (CatCh, internal ribosome entry site, β-galactosidase and an SV40 polyadenylation signal) was inserted into the ATG start site of the BAC Cdh5 gene (replacing the initiation codon of Cdh5 in exon 2). No other loc i on the BAC were altered. The 11.6 kbp BAC vector (pBACe3.6) contained a chloramphenicol selection cassette.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Research Tools
- Genetics Research
  - Tissue/Cell Markers: endothelial cells
  - Tissue/Cell Markers: multiple
  - Tissue/Cell Markers: transplantation marker for embryonic and adult tissue
  - Tissue/Cell Markers
- Diabetes and Obesity Research
  - lacZ
- Neurobiology Research
  - cell marker
- lacZ Expression
- Cardiovascular Research
- Developmental Biology Research
  - transplantation marker for embryonic and adult tissue
Neurobiology Research
- Optogenetic and Chemogenetic tools
  - Channelrhodopsin expressing strains
- Channel and Transporter Defects
- lacZ expression in neural tissue
Cardiovascular Research
- Vascular Defects
- Heart Abnormalities
- Other
Developmental Biology Research
- Internal/Organ Defects
  - brain
  - lung
  - heart
  - heart: vasculature
Internal/Organ Research
- Lung Defects
- Heart Abnormalities

Mammalian Phenotype Terms by Genotype

References

Additional - Tg(Cdh5-COP4*L132C,-lacZ)B28-2Mik related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Probe:Generic Cre Probe
- Standard PCR:Generic LacZ
- Standard PCR:Generic Cre Melt Curve Analysis
- Standard PCR:Generic LacZ Melt Curve Analysis
- Probe:Generic LacZ Probe
- QPCR:Generic LacZ QPCR Alternate 1
- Standard PCR:Tg(Cdh5-COP4*L132C,-lacZ)B28
- Genotyping resources and troubleshooting
Breeding Considerations

Mice hemizygous for the Cdh5-CatCh-IRES-lacZ transgene are viable and fertile with normal breeding, and no reported gross phenotypic abnormalities. When maintaining our live colony, hemizygous mice may be bred to wildtype (noncarrier) mice from the colony or to C57BL/6J inbred mice (Stock No. 000664).
To date (July 2019), it has not been attempted to make this strain homozygous.
- Additional Breeding and Husbandry Support
Mating System
- Hemizygote x Noncarrier
- Noncarrier x Hemizygote
Citation
When using the Cdh5-CatCh-IRES-lacZ , Cdh5^BAC-CatCh-IRES-lacZ mouse strain in a publication, please cite the originating article(s) and include JAX stock #033345 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Pricing & Availability

Cryo Recovery

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service/Product	Description	Price
	Hemizygous or non-carrier for Tg(Cdh5-COP4*L132C,-lacZ)B28-2Mik

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General Terms and Conditions

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Licensing Information

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Also Known As:Cdh5-CatCh-IRES-lacZ , Cdh5BAC-CatCh-IRES-lacZ

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Control Suggestions

Approximate Controls

Additional Information

Tg(Cdh5-COP4*L132C,-lacZ)B28-2Mik

Allele Symbol: Tg(Cdh5-COP4*L132C,-lacZ)B28-2Mik

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

References

Additional - Tg(Cdh5-COP4*L132C,-lacZ)B28-2Mik related

Genotyping Protocols

Breeding Considerations

Mating System

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Pricing

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Additional Use Restrictions Apply

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Also Known As:Cdh5-CatCh-IRES-lacZ , Cdh5^BAC-CatCh-IRES-lacZ