Strain Name:
B6.Cg-Hprttm360(Ple253-icre/ERT2)Ems/Mmjax
Stock Number:
037383-JAX
Citation ID:
RRID:MMRRC_037383-JAX
Major Collection:

Gene Information

Tg(Ovgp1-icre/ERT2)746Chok
Name: transgene insertion 746, Kathleen R Cho
Synonyms: Ovgp1-iCreERT2
Type: Allele
Species: Enterobacteria phage P1 (bacteriophage P1)
Chromosome: random insertion
Alteration at locus: Knock-In
Hprttm360a(Ple253-icre/ERT2)Ems
Name: hypoxanthine guanine phosphoribosyl transferase; targeted mutation 360a, Elizabeth M Simpson
Synonyms: Ple253-icre/frt/ERT2/frt, PITX3-creERT2, mEMS5929
Type: Allele
Species: Mus musculus (mouse)
Alteration at locus: Knock-In
Hprt
Name: hypoxanthine guanine phosphoribosyl transferase
Synonyms: Hprt1
Type: Gene
Species: Mus musculus (mouse)
Chromosome: X
Alteration at locus: Knock-In
NCBI: 15452
HGNC: HGNC:5157
Homologene: 56590
PITX3
Name: paired-like homeodomain transcription factor 3
Type: Gene
Species: Homo sapiens (human)
Chromosome: 10
Alteration at locus: Knock-In
Genetic Alterations
Ple253-icre/frt/ERT2/frt;mEMS5929 mice have the Ple253-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 paired-like homeodomain 3 (PITX3) 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.
Genotype Determination
Phenotype

Homozygous: Not yet evaluated

Hemizygous: Ple253-icre/frt/ERT2/frt;mEMS5929 mice have the Ple253-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 paired-like homeodomain 3 (PITX3) 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 (Diencephalon, midbrain, hindbrain)."

Strain Development
The Ple253-icre/frt/ERT2/frt transgene (pEMS2003) was designed with an HS4 insulator region, the 2484 bp Ple253 minipromoter (based on PITX3-B; derived from a portion of the endogenous promoter from the human paired-like homeodomain 3 (PITX3) 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. 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 ERT2 (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 Ple253-icre/frt/ERT2/frt transgene was targeted as a single copy knock-in to the Hprtb-m3 mutant locus on the X Chromosome via electroporation into mEMS4855.02 embryonic stem cells (described below). The resulting chimeric males (founder line mEMS5929) were bred to C57BL/6J females to establish the mutant colony. The donating investigator reports that Ple253-icre/frt/ERT2/frt;mEMS5929 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 MMRRC. When rederiving the live colony, an aliquot of frozen sperm is used to fertilize oocytes from C57BL/6J female mice; the resulting obligate heterozygous females are bred at least one generation to C57BL/6J mice to establish the live colony. The mEMS4855.02 embryonic stem (ES) cells are a male ES cell line made from an NE4.75F4 blastocyst from C57BL/6NTac-Aw-j/Aw-j Hprtb-m3/Y mice (created by four generations of breeding C57BL/6NTac-Aw-J together with C57BL/6NTac-Hprtb-m3). The parental lines were individually made by backcrossing C57BL/6J-Aw-J/J four generations onto C57BL/6NTac, and backcrossing B6.129P2-Hprtb-m3/J six generations onto C57BL/6NTac, respectively.
Donor
Simpson [Ph.D.], Elizabeth M.
Primary Reference

Some information may be available at the Pleiades Promoter and CanEuCre Projects website

Colony and Husbandry Information

Mice recovered from a cryo-archive will have health surveillance performed on recipient females. Health reports will be provided prior to shipment. If you require additional health status information, please email csmmrrc@jax.org .

Order Request Information

Limited quantities of breeder mice (recovered litter) are available from a cryoarchive; recovered litter usually available to ship in 3 to 4 months.

Cryopreserved material may be available upon request, please inquire to csmmrrc@jax.org for more information.

Distribution of this strain requires submission of the MMRRC Conditions of Use (COU). A link to the COU web form will be provided via email after an order has been placed; the form should be completed then or the email forwarded to your institutional official for completion.

The donor or their institution limits the distribution to non-profit institutions only.

Additional charges may apply for any special requests. Shipping costs are in addition to the basic distribution/resuscitation fees. Information on shipping costs and any additional charges will be provided by the supplying MMRRC facility.

Click button to Request this one strain. (Use the MMRRC Catalog Search to request more than one strain.)
MMRRC Item # Description Distribution Fee / Unit (US $)
*Shipping & Handling not included*
Units Notes
037383-JAX-SPERM Cryo-preserved spermatozoa $437.00 / Non-Profit Aliquot Approximate quantity3
037383-JAX-RESUS Litter recovered from cryo-archive $2,022.00 / Non-Profit Litter Recovered litter4; additional fees for any special requests.
Cryopreserved material may be available upon request, please inquire to csmmrrc@jax.org for more information.

1 The distribution fee covers the expense of rederiving mice from a live mouse; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.

2 An aliquot contains a sufficient number of embryos (in one or more vials or straws and based on the transfer success rate of the MMRRC facility) to transfer into one to three recipients. The MMRRC makes no guarantee concerning embryo transfer success experienced in the recipient investigator's laboratory. Neither gender nor genotype ratios are guaranteed.

3 An aliquot is one straw or vial with sufficient sperm to recover at least one litter of mice, as per provided protocols, when performed at the MMRRC facility. The MMRRC makes no guarantee concerning the success of these procedures when performed outside the MMRRC facilities.

4 The distribution fee covers the expense of resuscitating mice from the cryo-archive; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.

To request material from the MMRRC: Please fill out our on-line request form (accessible from the catalog search results page, or click the Request this Strain button in the fees section). If you have questions or need assistance completing this form, you may call Customer Service at (800) 910-2291 (in USA or Canada) or (530) 757-5710 (international calls). Before you call, please have with you: the MMRRC item number, quantity needed, Bill-to and Ship-to contact information.