Overview

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

Pvalb-T2A-dCre-D (Pvalb-2A-dCre-D) mice express a trimethoprim-inducible Cre recombinase directed to Pvalb-expressing cells by the endogenous promoter/enhancer elements of the parvalbumin locus. When induced, Cre recombinase activity is observed in scattered cells throughout the cortex, as well as restricted populations in the cerebellum, medulla, pons, pallidum, and thalamus.

Donating Investigator

Hongkui Zeng, Allen Institute for Brain Science

Genetic overview

Genetic Background	Generation

Pvalb^{tm5.1(cre/folA)Hze}

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

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Research Applications

Research Tools
Neurobiology Research
Cancer Research

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

The Pvalb-T2A-dCre-D targeted mutation (also called Pvalb-2A-dCre-D, Pvalb-2A-dCre-Δ or Pvalb-2A-dCre-Δhygro) has a viral 2A oligopeptide (T2A) that mediates ribosomal skipping and a destabilized Cre fusion gene (dCre) inserted in-frame at the 3' end of the parvalbumin (Pvalb) coding region. This is designed to have both endogenous gene and dCre expression directed to Pvalb-expressing cells by the endogenous promoter/enhancer elements.

The ecDHFR^R12Y/Y100I domain of dCre leads to proteosomal degradation of the entire fusion protein, resulting in little or no Cre recombinase activity. Administration of the DHFR inhibitor, trimethoprim (TMP), prevents degradation of the dCre fusion gene and results in Cre recombinase activity.

When Pvalb-T2A-dCre-D mice are bred with mice containing loxP-flanked sequences, TMP-stabilized Cre recombination will result in deletion of floxed sequences in the Pvalb-expressing cells of the double mutant offspring.

Specifically, the donating investigator reports that Pvalb-T2A-dCre-D mice have trimethoprim-inducible Cre recombinase expression (in situ hybridization) that is specific and efficient in scattered cells throughout the cortex, as well as restricted populations in the cerebellum, medulla, pons, pallidum, and thalamus.

In the absence of trimethoprim, these same regions exhibit significantly reduced Cre recombinase activity (highest expression in particular cells of the cerebellum and in the thalamic reticular nucleus). Heterozygous mice are viable and fertile with no gross physical or behavioral abnormalities. The donating investigator did not examine dCre activity in tissues other than brain, and did not attempt to generate homozygous mice to date (July 2013).

For characterization information, see images at the Allen Institute for Brain Science website (Pvalb-T2A-dCre images).

The dCre fusion gene (also called destabilized Cre, hDHFR/Cre or ecDHFR^R12Y/Y100I/Cre) is Cre recombinase fused at its N terminus to the first 159 amino acids of the Escherichia coli K-12 strain chromosomal dihydrofolate reductase gene (DHFR or folA) harboring the G67S mutation and modified to also include the R12Y/Y100I destabilizing domain mutations. The ecDHFR^R12Y/Y100I domain of dCre leads to proteosomal degradation of the entire fusion protein, resulting in little or no Cre recombinase activity. The donating investigator reports that administration of the high affinity DHFR inhibitor trimethoprim (TMP; at a concentration of 0.25-0.30 mg/g body weight) prevents degradation of the dCre fusion gene, resulting in Cre recombinase activity.

Development

The Pvalb-T2A-dCre-D targeted mutation (also called Pvalb-2A-dCre-D, Pvalb-2A-dCre-Δ or Pvalb-2A-dCre-Δhygro) was created in the laboratory of Dr. Hongkui Zeng (Allen Institute for Brain Science) to have a T2A sequence and a destabilized Cre fusion gene (dCre) 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-hDHFR/Cre" vector and a FLP-expressing plasmid to facilitate recombination.

Correctly re-targeted ES cells had (from 5' to 3')
a partial Pvalb intron 3 sequence containing an frt3 site,
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 dCre fusion gene (described below) that is in-frame with the Pvalb coding sequence,
the Pvalb 3' UTR sequence from exon 4,
an AttB site,
a PGK-hygromycin-SV40polyA cassette (with an mRNA splice donor-frt5 site-mRNA splice acceptor in the hygromycin gene),
and an AttP site.

The dCre fusion gene (also called destabilized Cre, hDHFR/Cre or ecDHFR^R12Y/Y100I/Cre) is Cre recombinase fused at its N terminus to the first 159 amino acids of the Escherichia coli K-12 strain chromosomal dihydrofolate reductase gene (DHFR or folA) harboring the G67S mutation and modified to also include the R12Y/Y100I destabilizing domain mutations.

Correctly re-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 sequences (PGK-5'hygro::mRNA splice donor::frt5::mRNA splice acceptor::3'hygro::SV40 polyA) and replace it with the recombined AttB/AttP site (AttL). The resulting Pvalb-T2A-dCre-D mice were bred with C57BL/6J wildtype mice for several generations (and the PhiC31 transgene was removed) prior to sending generation N6F1 males to The Jackson Laboratory Repository in 2013. Upon arrival, males were used to cryopreserve sperm. To establish the living Pvalb-T2A-dCre-D mouse colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J inbred females (Stock No. 000664).

Expression Data

Expressed Gene	cre/folA, cre/folA,
Site of Expression	With trimethoprim induction, cre expression (by (in situ hybridization detection) is efficient in scattered cells throughout the cortex, as well as restricted populations in the cerebellum, medulla, pons, pallidum, and thalamus.

Control Suggestions

Wild-type from the colony
000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Harris JA; Hirokawa KE; Sorensen SA; Gu H; Mills M; Ng LL; Bohn P; Mortrud M; Ouellette B; Kidney J; Smith KA; Dang C; Sunkin S; Bernard A; Oh SW; Madisen L; Zeng H. 2014. Anatomical characterization of Cre driver mice for neural circuit mapping and manipulation. Front Neural Circuits 8:76PubMed: 25071457 MGI: J:220523

View All References

Genetics

Pvalb^{tm5.1(cre/folA)Hze}

Allele Symbol: Pvalb^{tm5.1(cre/folA)Hze}

Allele Name	targeted mutation 5.1, Hongkui Zeng
Allele Type	Targeted (Recombinase-expressing, Inducible)
Allele Synonym(s)	Pvalb-2A-dCre; Pvalb-2A-dCre-DeltaHygrO; Pvalb-2A-hDHFR/Cre-D; Pvalb-2A-hDHFR/Cre-DeltaHygro
Gene Symbol and Name	Pvalb, parvalbumin
Gene Synonym(s)
Promoter	Pvalb, parvalbumin, mouse, laboratory
Expressed Gene	cre/folA, cre/folA,
Site of Expression	With trimethoprim induction, cre expression (by (in situ hybridization detection) is efficient in scattered cells throughout the cortex, as well as restricted populations in the cerebellum, medulla, pons, pallidum, and thalamus.
Strain of Origin	(129S6/SvEvTac x C57BL/6NCrl)F1
Chromosome	15
Molecular Note	ES cells previously targeted with a T2A-Cre vector immediately downstream of the parvalbumin translational STOP codon were re-targeted with a "T2A-hDHFR/Cre" vector and a FLP-expressing plasmid to facilitate recombination. Correctly re-targeted ES cells had (from 5' to 3') a partial Pvalb intron 3 sequence containing an frt3 site, a partial Pvalb exon 4 sequence up to (but not including) the endogenous stop codon, a T2A in-frame with the Pvalb coding sequence, a dCre fusion gene that is in-frame with the Pvalb codingsequence, the Pvalb 3' UTR sequence from exon 4, an AttB site, a PGK-hygromycin-SV40polyA cassette (with an mRNA spice donor-frt5 site-mRNA spice acceptor in the hygromycin gene), and an AttP site. The dCre fusion gene ( destabilized Cre, hDHFR/Cre or ecDHFR/Cre) is Cre recombinase fused at its N terminus to the first 159 amino acids of the E.coli K-12 strain chromosomal dihydrofolate reductase gene (DHFR or folA) with the G67S mutation and modified to include the R12Y/Y100I destabilizing domain mutations. Correctly re-targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were bred to PhiC31-expressing mice to delete the AttB/AttP-flanked sequences and replace them with the recombined AttB/AttP site (AttL). With trimethoprim induction, cre expression (by ISH detection) is efficient in scattered cells throughout the cortex, as well as restricted populations in thecerebellum, medulla, pons, pallidum, and thalamus.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Research Tools
- Neurobiology Research
  - cell marker
- Genetics Research
  - Tissue/Cell Markers: neurons
  - Mutagenesis and Transgenesis: Cre-lox System
  - Tissue/Cell Markers: Cre-lox System
  - Tissue/Cell Markers
  - Mutagenesis and Transgenesis
- Cre-lox System
  - Cre Recombinase Expression
  - Cre Recombinase Expression: Inducible
Neurobiology Research
- Cre-lox System
  - Cre Recombinase expression in neural tissue
Cancer Research
- Cre-lox System
  - Cre Recombinase expression: inducible

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^{tm5.1(cre/folA)Hze}/Pvalb⁺
involves: 129S6/SvEvTac * C57BL/6NCrl

normal phenotype

no abnormal phenotype detected
- heterozygotes are viable and fertile with no gross physical or behavioral abnormalities

References

Harris JA; Hirokawa KE; Sorensen SA; Gu H; Mills M; Ng LL; Bohn P; Mortrud M; Ouellette B; Kidney J; Smith KA; Dang C; Sunkin S; Bernard A; Oh SW; Madisen L; Zeng H. 2014. Anatomical characterization of Cre driver mice for neural circuit mapping and manipulation. Front Neural Circuits 8:76PubMed: 25071457 MGI: J:220523

Additional - Pvalb^{tm5.1(cre/folA)Hze} related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Pvalb
- 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 (July 2013).
- Additional Breeding and Husbandry Support
Citation
When using the Pvalb-T2A-dCre-D , Pvalb-2A-dCre-D mouse strain in a publication, please cite the originating article(s) and include JAX stock #022863 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/Product	Description	Price
	Heterozygous or Wildtype for Pvalb<tm5.1(cre/folA)Hze>

Related Products and Services

Frozen Mouse Embryo	B6.Cg-Pvalb<tm5.1(cre/folA)Hze>/J Frozen Embryo	$2595.00
Frozen Mouse Embryo	B6.Cg-Pvalb<tm5.1(cre/folA)Hze>/J Frozen Embryo	$2595.00
Frozen Mouse Embryo	B6.Cg-Pvalb<tm5.1(cre/folA)Hze>/J Frozen Embryo	$3373.50
Frozen Mouse Embryo	B6.Cg-Pvalb<tm5.1(cre/folA)Hze>/J Frozen Embryo	$3373.50

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

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Pvalbtm5.1(cre/folA)Hze

Allele Symbol: Pvalbtm5.1(cre/folA)Hze

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Pvalbtm5.1(cre/folA)Hze/Pvalb+involves: 129S6/SvEvTac * C57BL/6NCrl

normal phenotype

References

Additional - Pvalbtm5.1(cre/folA)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

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

Pvalb^{tm5.1(cre/folA)Hze}

Allele Symbol: Pvalb^{tm5.1(cre/folA)Hze}

Genotype: Pvalb^{tm5.1(cre/folA)Hze}/Pvalb⁺
involves: 129S6/SvEvTac * C57BL/6NCrl

Additional - Pvalb^{tm5.1(cre/folA)Hze} related