Overview

Also Known As: Pvalb-2A-FlpE-D , Pvalb-T2A-FlpE-D

Pvalb-2A-FlpE-D (Pvalb-T2A-FlpE-D) mice have both endogenous Pvalb gene and sequence-enhanced Flp recombinase (FlpE) gene expression directed to Pvalb-expressing cells by the endogenous promoter/enhancer elements of the parvalbumin locus. These mice are a tool that allows deletion of frt-flanked sequences in Pvalb-expressing cells/tissues.

Donating Investigator

Hongkui Zeng, Allen Institute for Brain Science

Genetic overview

Genetic Background	Generation

Pvalb^{tm2.1(flpe)Hze}

Allele Type	Gene Symbol	Gene Name
Targeted (Recombinase-expressing)	Pvalb	parvalbumin

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Research Applications

Research Tools

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

The Pvalb-2A-FlpE-D (Pvalb-T2A-FlpE-D) targeted mutation has a viral 2A oligopeptide (T2A) that mediates ribosomal skipping and a sequence-enhanced FlpE recombinase gene inserted in-frame at the 3' end of the parvalbumin (Pvalb) coding region.

As such, Pvalb-2A-FlpE-D mice have both endogenous Pvalb gene and FlpE expression directed to Pvalb-expressing cells by the endogenous promoter/enhancer elements of the parvalbumin locus. When Pvalb-2A-FlpE-D mice are bred with mice containing frt-flanked sequences, FlpE-mediated recombination will result in deletion of frt-flanked sequences in the Pvalb-expressing cells of the double mutant offspring.

Specifically, the donating investigator reports heterozygous mice have an FlpE recombinase expression pattern in the brain that is very similar to endogenous Pvalb expression (by in situ hybridization using a FlpE-specific probe). The donating investigator did not examine FlpE activity in tissues other than brain, and did not attempt to generate homozygous mice to date (January 2013).

Mice heterozygous for the Pvalb-2A-FlpE-D allele are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities.

Development

Pvalb-2A-FlpE-D (Pvalb-T2A-FlpE-D) mice were created in the laboratory of Dr. Hongkui Zeng (Allen Institute for Brain Science) to have a T2A sequence and a sequence-enhanced FlpE recombinase gene inserted in-frame at the 3' end of the parvalbumin (Pvalb) coding region on chromosome 15. The specific details are below.

(129S6/SvEvTac x C57BL/6)F1-derived G4 embryonic stem (ES) cells, already targeted with a T2A-Cre vector inserted in-frame at the 3' end of the Pvalb coding region (see Stock No. 012358), were re-targeted with a "T2A-FlpE" vector and a FLP-expressing plasmid to facilitate recombination. Correctly targeted ES cells had (from 5' to 3') a three amino acid linker sequence, an frt3 site within Pvalb intron 3, a partial Pvalb exon 4 sequence up to (but not including) the endogenous stop codon, a viral 2A oligopeptide (T2A; mediates ribosomal skipping) that is in-frame with the Pvalb coding sequence, a sequence-enhanced FlpE recombinase gene, the Pvalb 3' UTR sequence from exon 4, an attB site, a PGK-hygromycin-SV40polyA cassette (with an RNA splice donor-frt5 site-RNA splice acceptor in the hygromycin gene), and an attP site.

Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were bred to PhiC31-expressing mice (C57BL/6J congenic background; see Stock No. 007743) to delete the attB/attP-flanked PGK-hygromycin-polyA cassette and replace it with the recombined attB/attP site (attL). The resulting Pvalb-2A-FlpE-D mice were bred with C57BL/6J wildtype mice for at least seven additional generations (and the PhiC31 transgene was removed) prior to sending to The Jackson Laboratory Repository in 2013.

Upon arrival, sperm as cryopreserved. To establish the living Pvalb-2A-FlpE-D mouse colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J inbred females (Stock No. 000664).

Control Suggestions

Wild-type from the colony
000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Madisen L; Zwingman TA; Sunkin SM; Oh SW; Zariwala HA; Gu H; Ng LL; Palmiter RD; Hawrylycz MJ; Jones AR; Lein ES; Zeng H. 2010. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat Neurosci 13(1):133-40PubMed: 20023653 MGI: J:155793
Madisen L; Garner AR; Shimaoka D; Chuong AS; Klapoetke NC; Li L; van der Bourg A; Niino Y; Egolf L; Monetti C; Gu H; Mills M; Cheng A; Tasic B; Nguyen TN; Sunkin SM; Benucci A; Nagy A; Miyawaki A; Helmchen F; Empson RM; Knopfel T; Boyden ES; Reid RC; Carandini M; Zeng H. 2015. Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance. Neuron 85(5):942-58PubMed: 25741722 MGI: J:219930

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Genetics

Pvalb^{tm2.1(flpe)Hze}

Allele Symbol: Pvalb^{tm2.1(flpe)Hze}

Allele Name	targeted mutation 2.1, Hongkui Zeng
Allele Type	Targeted (Recombinase-expressing)
Allele Synonym(s)	targeted mutation 2.1, Hongkui Zeng; Pvalb^{tm2.1(flpe)Hze}
Gene Symbol and Name	Pvalb, parvalbumin
Gene Synonym(s)	Pva; PV; Pva; Parv; D22S749; PALB1
Promoter	Pvalb, parvalbumin, mouse, laboratory
Site of Expression	Heterozygous mice have an FlpE recombinase expression pattern in the brain that is very similar to endogenous Pvalb expression.
Strain of Origin	(129S6/SvEvTac x C57BL/6NCrl)F1
Chromosome	15
Molecular Note	Embryonic stem (ES) cells, previously targeted with a T2A-Cre vector immediately downstream of the parvalbumin translational STOP codon were re-targeted with a T2A-FLPe (enhanced flippase) vector and a FLP-expressing plasmid to facilitate recombination. Correctly targeted ES cells had (from 5' to 3') a three amino acid linker sequence, an frt3 site within Pvalb intron 3, a partial Pvalb exon 4 sequence up to (but not including) the endogenous stop codon, a T2A sequence that is in-frame with the Pvalb coding sequence, a sequence-enhanced flippase gene (FLPe), the Pvalb 3' UTR sequence from exon 4, an attB site, a PGK-hygromycin-SV40polyA cassette and an attP site. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were bred to PhiC31-expressing mice to delete the attB/attP-flanked PGK cassette. The resulting mice were bred with C57BL/6J wildtype mice for at least two additional generations (and the PhiC31 transgene was removed).

Disease/Phenotype

Disease Terms

Research Areas By Genotype

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

Research Tools
- FLP-FRT System
  - FLP Recombinase Expression
- Genetics Research
  - Mutagenesis and Transgenesis: FLP recombinase
  - Tissue/Cell Markers
  - Mutagenesis and Transgenesis
- Neurobiology Research
  - cell marker

Mammalian Phenotype Terms by Genotype

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

Genotype: Pvalb^{tm2.1(flpe)Hze}/Pvalb⁺
involves: 129S6/SvEvTac * C57BL/6J * C57BL/6NCr

mammalian phenotype

no phenotypic analysis
- Normal - no analysis is available

References

Madisen L; Zwingman TA; Sunkin SM; Oh SW; Zariwala HA; Gu H; Ng LL; Palmiter RD; Hawrylycz MJ; Jones AR; Lein ES; Zeng H. 2010. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat Neurosci 13(1):133-40PubMed: 20023653 MGI: J:155793
Madisen L; Garner AR; Shimaoka D; Chuong AS; Klapoetke NC; Li L; van der Bourg A; Niino Y; Egolf L; Monetti C; Gu H; Mills M; Cheng A; Tasic B; Nguyen TN; Sunkin SM; Benucci A; Nagy A; Miyawaki A; Helmchen F; Empson RM; Knopfel T; Boyden ES; Reid RC; Carandini M; Zeng H. 2015. Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance. Neuron 85(5):942-58PubMed: 25741722 MGI: J:219930

Additional - Pvalb^{tm2.1(flpe)Hze} related

Technical Support

Contact Technical Support

Genotyping Protocols
- High Resolution Melting: Tg(ACTFLPe)
- SEPARATED MELT: Pvalb^{tm2.1(FLPe)Hze}alternate3
- Probe: Generic FLP
- Standard PCR: Tg(ACTFLPe)
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, heterozygous mice may be bred to wildtype mice from the colony or to C57BL/6J inbred mice (Stock No. 000664). The phenotype of homozygous mice has not yet been determined (January 2013).
- Additional Breeding and Husbandry Support
Citation
When using the Pvalb-2A-FlpE-D , Pvalb-T2A-FlpE-D mouse strain in a publication, please cite the originating article(s) and include JAX stock #021191 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 - Not-For-Profit & Academic Pricing

Service	Genotype	Price
	Heterozygous or wildtype for Pvalb<tm2.1(FLPe)Hze>/

We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.

Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation.

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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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Strain(s) not available to companies or for-profit entities.

Licensing Information

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Also Known As: Pvalb-2A-FlpE-D , Pvalb-T2A-FlpE-D

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Control Suggestions

Additional Information

Selected References

Pvalbtm2.1(flpe)Hze

Allele Symbol: Pvalbtm2.1(flpe)Hze

Disease Terms

Research Areas By Genotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Pvalbtm2.1(flpe)Hze/Pvalb+involves: 129S6/SvEvTac * C57BL/6J * C57BL/6NCr

mammalian phenotype

References

Additional - Pvalbtm2.1(flpe)Hze 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 - Not-For-Profit & Academic 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

Pvalb^{tm2.1(flpe)Hze}

Allele Symbol: Pvalb^{tm2.1(flpe)Hze}

Genotype: Pvalb^{tm2.1(flpe)Hze}/Pvalb⁺
involves: 129S6/SvEvTac * C57BL/6J * C57BL/6NCr

Additional - Pvalb^{tm2.1(flpe)Hze} related