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Strain Name:
B6.129-Rr27tm1Kpfe/Mmjax
Stock Number:
071834-JAX
Citation ID:
RRID:MMRRC_071834-JAX
Other Names:
H19-ICRFlox

Strain Information

Rr27tm1Kpfe
Name: regulatory region 27; targeted mutation 1, Karl Pfeifer
Synonyms: DMRflox, H19tm1Kpfe
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 7
Alteration at locus: Conditional
Rr27
Name: regulatory region 27
Synonyms: IC1, ICR, DMD
Type: Regulatory Region
Species: Mus musculus (mouse)
Chromosome: 7
Alteration at locus: Conditional
Genetic Alterations
The DMRflox allele allows cre-mediated deletion of a 5.7kbp region, containing the differentially methylated domain (DMD or Rr27), upstream of the H19 start site. This DMD coordinates the reciprocal expression of H19 and Igf2 by controlling access to shared enhancers. These mice may be used to generate temporal and tissue-specific deletions of this imprinting control region for studying the chromatin interactions and epigenetics involved in H19/Igf2 imprinting.
Genotype Determination
ES Cell Line
R1 derived from (129X1/SvJ x 129S1/Sv)F1-Kitl+
Phenotype

Imprinted expression of the H19 and Igf2 loci is dependent upon a differentially methylated domain (DMD or Rr27) that coordinates the reciprocal maternal/paternal expression of H19 and Igf2 by controlling access to shared enhancers. Specifically, on the maternal allele, the DMD acts as a methylation-sensitive insulator that results in H19 expression and Igf2 inactivation. The four CpG-rich regions in the DMD bind CCCTC-binding factor (CTCF) on the hypomethylated maternal allele, subsequently blocking the Igf2 gene from the enhancers shared by H19 and Igf2. On the paternal allele, the hypermethylation of DMD concomitantly methylates H19, resulting in H19 inactivation and Igf2 expression. The DMRflox mice have loxP sites flanking the H19/Igf2 imprinting control region, at -6.5 to -0.8kb upstream of the H19 major start site.

Mice homozygous for the DMRflox allele are viable and fertile. After Cre recombinase-deletion of the floxed DMD region, the resulting mice exhibit tissue-specific disruption of imprinted expression of H19 and Igf2. The donating laboratory notes that upon Cre-mediated recombination on the maternal chromosome there is a phenotype that mimics Beckwith Wiedemann syndrome. Maternal Igf2 expression is activated so that overall there is 2X expression of Igf2 and maternal H19 expression is reduced by 4-50-fold in a tissue dependent manner. Together, these expression changes result in a series of phenotypes including fetal overgrowth, cancer susceptibility, and cardiac insufficiency in aged mice.

These DMRflox mice may be useful for studying the chromatin interactions and epigenetics involved in H19/Igf2 imprinting.

MeSH Terms
  • Alleles
  • Animals
  • Attachment Sites, Microbiological/genetics
  • Chromosomes/genetics
  • CpG Islands/genetics
  • DNA Methylation
  • Female
  • Gametogenesis/genetics
  • Gene Expression Regulation, Developmental/genetics
  • Gene Silencing
  • Genomic Imprinting/genetics
  • Insulin-Like Growth Factor II/genetics
  • Integrases/genetics
  • Integrases/metabolism
  • Male
  • Mice
  • Models, Genetic
  • Muscle Proteins/genetics
  • Mutagenesis, Insertional/genetics
  • Promoter Regions, Genetic/genetics
  • RNA, Long Noncoding
  • RNA, Messenger/genetics
  • RNA, Messenger/metabolism
  • RNA, Untranslated
  • Regulatory Sequences, Nucleic Acid/genetics
  • Sequence Deletion/genetics
  • Time Factors
  • Transcription, Genetic/genetics
  • Viral Proteins
Strain GQC Summary
Gene Specific Genotyping:

To request gene-specific and other genotyping services for a strain, please contact the distribution MMRRC Center for more information.

Background Genetic Quality:

The MMRRC has developed a Genetic Quality Control pipeline using the MiniMUGA array to provide additional information to identify and validate genetic backgrounds of MMRRC strains. For more information on whether genetic background data is available, please contact MMRRC_GeneticQC@med.unc.edu. Note: that MiniMUGA genetic background data is not available on all strains, but can be ordered if desired.

Suggested Control Mice
Littermates of all relevant genotypes.
  • Cardiovascular
  • Developmental Biology
  • Models for Human Disease
Donor
Karl Pfeifer, National Institutes of Health (NIH), NICHD.
Primary Reference

Srivastava M, Hsieh S, Grinberg A, Williams-Simons L, Huang SP, Pfeifer K. H19 and Igf2 monoallelic expression is regulated in two distinct ways by a shared cis acting regulatory region upstream of H19. Genes Dev. 2000 May 15;14(10):1186-95. (Medline PMID: 10817754)

Strain Development
The DMRflox allele was designed to have loxP sites flanking -6.5 to -0.8kb upstream of the H19 major start within the regulatory region 27 (Rr27; a differentially methylated domain [DMD]) on chromosome 7. Specifically, the targeting vector contained sequences extending from -11kb EcoRI to +6.5kb XbaI relative to the H19 transcriptional start site. The construct also contained a neomycin-resistance gene flanked by loxP sites inserted at the -7.0kb HindIII site and another loxP site inserted at the -0.8kb XbaI site. The targeting vector was electroporated into (129X1/SvJ x 129S1/Sv)F1-Kitl+-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were then transiently transfected with a Cre recombinase-expressing plasmid. The resulting ES cells with the DMRflox genotype (retaining the floxed DMD with the neomycin cassette deleted) were injected into recipient blastocysts. Chimeric mice were bred with C57BL/6J mice to establish germline transmission; the donating laboratory further backcrossed the DMRflox mice to C57BL/6J mice for at least 8 generations. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. One of the 43 markers throughout the genome were segregating with 129. Also, all 4 of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.


Disclaimer: If MMRRC Strain Genetic Quality Control (GQC; based on MiniMUGA genotyping and analysis) has been completed for this strain, the information might differ from the genetic background information provided by the submitter. MiniMUGA genetic analysis is done on a strain’s tissue samples taken when archived by or ordered from the assigned MMRRC Center.

Colony and Husbandry Information

Cryo-recovered strains distributed by the MMRRC at JAX are shipped to the customer from the Pathogen & Opportunistic-Free Animal Room G200 - see https://www.jax.org/jax-mice-and-services/customer-support/customer-service/animal-health/health-status-reports.

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.
Coat Color
Black
Eye
Black
MMRRC Breeding System
Backcross
Generation
8
Overall Breeding Performance
Good
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Yes Yes
Heterozygotes are fertile: Yes Yes
Age Reproductive Decline: Greater than 12 months Undetermined
Average litter size
4 to 6
Recommended wean age
3 Weeks
Average Pups Weaned
4 to 6

Order 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.

A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Nathan Whitman.

A Commercial License Agreement from the Submitter is required for for-profit entities to use this strain. For more information, please contact Nathan Whitman

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.

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
071834-JAX-SPERM Cryo-preserved spermatozoa $459.00 / $459.00
Non-Profit / For-Profit
Aliquot Approximate quantity3
071834-JAX-RESUS Litter recovered from cryo-archive $2,123.00 / $2,123.00
Non-Profit / For-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.