Overview

Also Known As:Ple155-icre/frt/ERT2/frt;mEMS5985, C57BL/6-Hprt^{tm347(Ple155-icre/ERT2)Ems}/Mmjax, PCP2-creERT2

Ple155-icre/frt/ERT2/frt;mEMS5985 mice have the Ple155-icre/frt/ERT2/frt transgene targeted as a single copy "knock-in" into the upstream region of the hypoxanthine guanine phosphoribosyl transferase (Hprt) locus on the X Chromosome. This is designed to allow specific portions from the promoter/enhancer/regulatory regions of the human purkinje cell protein 2 (PCP2) locus to direct expression of a tamoxifen-inducible, improved Cre recombinase (icre/ERT2). In these mice, the icre/ERT2 estrogen receptor sequences are flanked with frt sites.

Donating Investigator

Elizabeth M Simpson, Centre for Molecular Medicine & Therapeutics, University of British Columbia

Genetic overview

Genetic Background	Generation

Hprt^{tm347a(Ple155-icre/ERT2)Ems}

Allele Type	Gene Symbol	Gene Name
Targeted (Recombinase-expressing, Inducible)	Hprt	hypoxanthine guanine phosphoribosyl transferase

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Research Applications

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Details

Detailed Description

Ple155-icre/frt/ERT2/frt;mEMS5985 mice have the Ple155-icre/frt/ERT2/frt transgene targeted as a single copy "knock-in" into the upstream region of the hypoxanthine guanine phosphoribosyl transferase (Hprt) locus on the X Chromosome. Heterozygous females and hemizygous males are viable and fertile, with specific portions from the promoter/enhancer/regulatory regions of the human purkinje cell protein 2 (PCP2) locus directing expression of the tamoxifen-inducible, improved Cre recombinase (icre/ERT2). In these mice, the icre/ERT2 estrogen receptor sequences are flanked with frt sites. The donating investigator reports RT PCR expression as "Positive (Eyes, Diencephalon, Cerebellum)." The phenotype of homozygous mice has not been evaluated to date (January 2014).

The iCre/ER^T2 fusion protein used here consists of a codon-improved Cre recombinase fused to a G400V/M543A/L544A triple mutant form of the human estrogen receptor which does not bind its natural ligand (17β-estradiol) at physiological concentrations but will bind the synthetic estrogen receptor ligands 4-hydroxytamoxifen (OHT or tamoxifen) and, with lesser sensitivity, ICI 182780. Restricted to the cytoplasm, iCre/ER^T2 can only gain access to the nuclear compartment after exposure to tamoxifen. To counteract the mixed estrogen agonist effects of tamoxifen injections, which can result in late fetal abortions in pregnant mice, progesterone may be coadministered. The iCre/ER^T2 fusion protein used here has also been modified to have F3-frt sites flanking the estrogen receptor sequences.

Development

Ple155-icre/frt/ERT2/frt;mEMS5985 mice were created by the CanEuCre and Pleiades Promoter Project (Centre for Molecular Medicine and Therapeutics, University of British Columbia). These mice have the Ple155-icre/frt/ERT2/frt transgene targeted as a single copy "knock-in" into the upstream region of the hypoxanthine guanine phosphoribosyl transferase (Hprt) locus on the X Chromosome. As such, specific regions from the promoter/enhancer/regulatory sequences of the human purkinje cell protein 2 (PCP2) locus are designed to direct expression of the tamoxifen-inducible, improved Cre recombinase (icre/ERT2). In these mice, the icre/ERT2 estrogen receptor sequences are flanked with frt sites. More specific details are described below.

The Ple155-icre/frt/ERT2/frt transgene (pEMS2004) was designed with an HS4 insulator region, the 1652 bp Ple155 minipromoter (based on PCP2-B; derived from a portion of the endogenous promoter (Prom 1 and Prom 2) from the human purkinje cell protein 2 (PCP2) locus), a chimeric intron cassette, the iCre/ER^T2 fusion gene (with F3-frt sites flanking the ER^T2 sequences), the mut6-WPRE element, an SV40 early polyA signal, an additional HS4 insulator region, a human HPRT complementary sequence (containing exon 1, intron 1, exon 2, and part of intron 2), a mouse 3' Hprt homology arm, an I-Sce linearization cut site, and a mouse 5' Hprt homology arm. The iCre/ERT2 fusion gene used here has an optimized variant of Cre recombinase (iCre; improved with mammalian codon usage, removed putative cryptic splice sites, altered stop codon, and reduced CpG content to limit the chances of epigenetic silencing in mammals) fused to ER^T2 (a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand binding domain); additionally modified to have F3-frt sites flanking the estrogen receptor sequences. The Ple155-icre/frt/ERT2/frt transgene was targeted as a single copy knock-in to the Hprt^b-m3 mutant locus on the X Chromosome via electroporation into mEMS4855.02 embryonic stem cells (described below). The resulting chimeric males (founder line mEMS5985) were bred to C57BL/6J females to establish the mutant colony. The donating investigator reports that Ple155-icre/frt/ERT2/frt;mEMS5985 mutant animals were backcrossed to C57BL/6J mice for 2 generations, and then sperm was frozen from black hemizygous males at the University of British Columbia. In 2013-2014, frozen sperm was sent to the Mutant Mouse Regional Resource Center node at The Jackson Laboratory (MMRRC-JAX). When rederiving the live colony, an aliquot of frozen sperm is used to fertilize oocytes from C57BL/6J female mice (Stock No. 000664); the resulting obligate heterozygous females are bred at least one generation to C57BL/6J mice to establish the live colony.

mEMS4855.02 embryonic stem (ES) cells are a male ES cell line made from an NE4.75F4 blastocyst from C57BL/6NTac-A^w-j/A^w-j Hprt^b-m3/Y mice (created by four generations of breeding C57BL/6NTac-A^w-J together with C57BL/6NTac-Hprt^b-m3). The parental lines were individually made by backcrossing C57BL/6J-A^w-J/J (Stock No. 000051) four generations onto C57BL/6NTac, and backcrossing B6.129P2-Hprt^b-m3/J (Stock No. 002171) six generations onto C57BL/6NTac, respectively.

Expression Data

Expressed Gene	cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of Expression

Control Suggestions

Approximate Controls

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Korecki AJ; Hickmott JW; Lam SL; Dreolini L; Mathelier A; Baker O; Kuehne C; Bonaguro RJ; Smith J; Tan CV; Zhou M; Goldowitz D; Deussing JM; Stewart AF; Wasserman WW; Holt RA; Simpson EM. 2019. Twenty-Seven Tamoxifen-Inducible iCre-Driver Mouse Strains for Eye and Brain; Including Seventeen Carrying a New Inducible-First Constitutive-Ready Allele. GeneticsPubMed: 30765420 MGI: J:270567

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Genetics

Hprt^{tm347a(Ple155-icre/ERT2)Ems}

Allele Symbol: Hprt^{tm347a(Ple155-icre/ERT2)Ems}

Allele Name	targeted mutation 347a, Elizabeth M Simpson
Allele Type	Targeted (Recombinase-expressing, Inducible)
Allele Synonym(s)	PCP2-creERT2; Ple155-icre/frt/ERT2/frt;mEMS5985
Gene Symbol and Name	Hprt, hypoxanthine guanine phosphoribosyl transferase
Gene Synonym(s)
Expressed Gene	cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene,
Strain of Origin	B6N.Cg-A^w-J Hprt^b-m3
Chromosome	X
Molecular Note	The transgene was designed with an HS4 insulator region, the 1652 bp Ple155 minipromoter (based on PCP2-B; derived from a portion of the endogenous promoter (Prom 1 and Prom 2) from the human purkinje cell protein 2 (PCP2) locus), a chimeric intron cassette, the iCre/ERT2 fusion gene (with F3-frt sites flanking the ERT2 sequences), the mut6-WPRE element, an SV40 early polyA signal, an additional HS4 insulator region, a human HPRT complementary sequence (containing exon 1, intron 1, exon 2, and part of intron 2), a mouse 3' Hprt homology arm, an I-Sce linearization cut site, and a mouse 5' Hprt homology arm. T

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

The following phenotype relates to a compound genotype created using this strain

Genotype: Hprt^{tm347a(Ple155-icre/ERT2)Ems}/Y
B6.Cg-Hprt

normal phenotype

no abnormal phenotype detected
- heterozygous females and hemizygous males are viable and fertile

References

Korecki AJ; Hickmott JW; Lam SL; Dreolini L; Mathelier A; Baker O; Kuehne C; Bonaguro RJ; Smith J; Tan CV; Zhou M; Goldowitz D; Deussing JM; Stewart AF; Wasserman WW; Holt RA; Simpson EM. 2019. Twenty-Seven Tamoxifen-Inducible iCre-Driver Mouse Strains for Eye and Brain; Including Seventeen Carrying a New Inducible-First Constitutive-Ready Allele. GeneticsPubMed: 30765420 MGI: J:270567

Additional - Hprt^{tm347a(Ple155-icre/ERT2)Ems} related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Generic iCre
- Probe:Generic iCre Probe
- Genotyping resources and troubleshooting
Breeding Considerations

The targeted mutation in on the X chromosome. When maintaining a live colony, heterozygous females may be bred with wildtype males from the colony or with C57BL/6J inbred males (Stock No. 000664). Alternatively, wildtype females from the colony or C57BL/6J inbred females may be bred with hemizygous males. Homozygous females and hemizygous males are expected to be viable and fertile. The expected coat color is black.
- Additional Breeding and Husbandry Support
Citation
When using the B6.Cg-Hprt^{tm347a(Ple155-icre/ERT2)Ems}/Mmjax mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #037077 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

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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Also Known As:Ple155-icre/frt/ERT2/frt;mEMS5985, C57BL/6-Hprttm347(Ple155-icre/ERT2)Ems/Mmjax, PCP2-creERT2

Donating Investigator

Genetic overview

Research Applications

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Hprttm347a(Ple155-icre/ERT2)Ems

Allele Symbol: Hprttm347a(Ple155-icre/ERT2)Ems

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Hprttm347a(Ple155-icre/ERT2)Ems/YB6.Cg-Hprt

normal phenotype

References

Additional - Hprttm347a(Ple155-icre/ERT2)Ems related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

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

The Jackson Laboratory's Genotype Promise

Terms Of Use

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:Ple155-icre/frt/ERT2/frt;mEMS5985, C57BL/6-Hprt^{tm347(Ple155-icre/ERT2)Ems}/Mmjax, PCP2-creERT2

Hprt^{tm347a(Ple155-icre/ERT2)Ems}

Allele Symbol: Hprt^{tm347a(Ple155-icre/ERT2)Ems}

Genotype: Hprt^{tm347a(Ple155-icre/ERT2)Ems}/Y
B6.Cg-Hprt

Additional - Hprt^{tm347a(Ple155-icre/ERT2)Ems} related