Overview

These Shh::gfp mice harbor a targeted mutation of the Sonic hedgehog (Shh) locus that secretes, after normal Shh processing, a bioactive GFP-tagged Shh signaling ligand (N-Shh::GFPp). These Shh::gfp may be useful to directly visualize the function of Shh ligand in an in vivo context, such as during neural tube patterning.

Donating Investigator

Andrew P McMahon, University of Southern California

Genetic overview

Genetic Background	Generation

Shh^tm6Amc

Allele Type	Gene Symbol	Gene Name
Targeted (Reporter, Null/Knockout)	Shh	sonic hedgehog

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Research Applications

Developmental Biology Research
Neurobiology Research
Research Tools

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

While mice heterozygous for the Shh::gfp allele are viable, fertile, and indistinguishable from wild-type littermates, homozygotes are stillborn and show developmental defects consistent with reduced Sonic Hedgehog (Shh) signaling. The Shh::gfp mutation has GFP inserted into the endogenous Shh processing site (and adds a new processing site after the GFP). Thus normal Shh processing leads to secretion of the GFP-tagged Shh signaling ligand (N-Shh::GFPp) instead of wild-type Shh; with N-Shh::GFPp retaining both GFP and lipid modifications post-processing. Biochemical and cellular analysis indicates that Shh::GFP undergoes correct processing to produce active, bi-lipidated signaling peptides. Shh::GFP processing is, however, less efficient and results in reduced levels of Shh::GFP compared with wild-type Shh protein. These Shh::gfp mice produce bioactive, fluorescently labeled Shh from the endogenous Shh locus and may be useful to directly visualize the function of Shh ligand in an in vivo context, such as during neural tube patterning. In addition, these mice may also be useful in conjunction with other Sonic Hedgehog (Shh) mutant strains including Shh knockout mice (Stock No. 003318), Shh-floxed mice (Stock No. 004293), Shh-GFP/cre mice (Stock No. 005622), and Shh-creERT2 mice (Stock No. 005623).

Development

A targeting vector was designed to replace the endogenous intein cleavage-cholesterol attachment site (Shh processing site) in exon 3 of the targeted gene with a Green Fluorescent Protein (GFP) followed by a new intein cleavage-cholesterol attachment site (Shh processing site), as well as insert a frt-flanked PGK-Neo cassette in intron 2. The donating investigator reports that this construct was microinjected into 129X1/SvJ-derived AV3 embryonic stem (ES) cells, correctly targeted ES cells were injected into recipient blastocysts, and chimeric mice were bred to C57BL/6 inbred mice to generate Shh::gfp mice. These Shh::gfp mice were reported to have been bred with C57BL/6 mice for a small number generations prior to sending to The Jackson Laboratory Repository in 2008 (see SNP notes below). Upon arrival, these mice were bred to C57BL/6J inbred mice (Stock No. 000664) to establish the colony. Over the next two years, the colony was backcrossed to C57BL/6J and/or wildtype mice from the colony for several additional generations. By March 2011, the colony was found to be ~96% C57BL/6 genetic background by SNP analysis (see SNP notes below).

In 2009, a 27 SNP (single nucleotide polymorphism) panel analysis performed on the rederived living colony at The Jackson Laboratory Repository. This revealed 16 markers (across several different chromosomes) that were not C57BL/6 allele-type. These data suggest the strain was sent to The Jackson Laboratory Repository with multiple sources of genetic background contamination (more than 129 genetic background).

In 2011, a 32 SNP (single nucleotide polymorphism) panel analysis, with markers covering all 19 chromosomes and the X chromosome, was performed on the colony at The Jackson Laboratory Repository. This revealed one marker on chromosome 1 (~89.1 Mbp) that was not fixed for C57BL/6 allele-type (e.g.: still segregating for 129X1 allele-type).

Expression Data

Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	early neuronal precursors

Selected References

Chamberlain CE; Jeong J; Guo C; Allen BL; McMahon AP. 2008. Notochord-derived Shh concentrates in close association with the apically positioned basal body in neural target cells and forms a dynamic gradient during neural patterning. Development 135(6):1097-106PubMed: 18272593 MGI: J:132152

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Genetics

Shh^tm6Amc

Allele Symbol: Shh^tm6Amc

Allele Name	targeted mutation 6, Andrew P McMahon
Allele Type	Targeted (Reporter, Null/Knockout)
Allele Synonym(s)	Shh::GFP
Gene Symbol and Name	Shh, sonic hedgehog
Gene Synonym(s)
Expressed Gene	GFP, Green Fluorescent Protein,
Site of Expression	early neuronal precursors
Strain of Origin	Not Specified
Chromosome	5
Molecular Note	A GFP cassette was inserted into exon 3 and an frt-flanked neo cassette was inserted between exon 2 and 3. The presence of the fusion protein and the absence of the endogenous product was confirmed by western blot analysis on E9.5 embryo extracts. This allele produces functional albeit inefficient protein product.
Mutations Made By	Andrew McMahon, University of Southern California

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Developmental Biology Research
- Embryonic Lethality (Homozygous)
- Limb Patterning Defects
- Skeletal Defects
- Neural Tube Defects
Neurobiology Research
- Fluorescent protein expression in neural tissue
Research Tools
- Fluorescent Proteins

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Shh^tm6Amc/Shh^tm6Amc
Not Specified

mortality/aging

lethality throughout fetal growth and development, incomplete penetrance
- homozygous mice are still born and show defects consistent with reduced Shh signaling
- Normal - neural tube at E10.5 show all Shh-dependent neural progenitor domains resulting in normal induction of floor plate

References

Chamberlain CE; Jeong J; Guo C; Allen BL; McMahon AP. 2008. Notochord-derived Shh concentrates in close association with the apically positioned basal body in neural target cells and forms a dynamic gradient during neural patterning. Development 135(6):1097-106PubMed: 18272593 MGI: J:132152

Additional - Shh^tm6Amc related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Shh
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, heterozygous mice may be bred together or to wildtype littermates.
- Additional Breeding and Husbandry Support
Citation
When using the B6.129X1(Cg)-Shh^tm6Amc/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #008466 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
	Heterozygous or Wild-type for Shh<tm6Amc>

Related Products and Services

Frozen Mouse Embryo	B6.129X1(Cg)-Shh<tm6Amc>/J Frozen Embryo	$2595.00
Frozen Mouse Embryo	B6.129X1(Cg)-Shh<tm6Amc>/J Frozen Embryo	$2595.00
Frozen Mouse Embryo	B6.129X1(Cg)-Shh<tm6Amc>/J Frozen Embryo	$3373.50
Frozen Mouse Embryo	B6.129X1(Cg)-Shh<tm6Amc>/J Frozen Embryo	$3373.50

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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. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.

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Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Selected References

Shhtm6Amc

Allele Symbol: Shhtm6Amc

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Shhtm6Amc/Shhtm6AmcNot Specified

mortality/aging

References

Additional - Shhtm6Amc related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

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

Live Mouse

Cryorecovery - Pricing

Related Products and Services

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

Shh^tm6Amc

Allele Symbol: Shh^tm6Amc

Genotype: Shh^tm6Amc/Shh^tm6Amc
Not Specified

Additional - Shh^tm6Amc related