Strain Name:
FVB/NCrl-Rr43em1Axvi Rr44em2Axvi/Mmucd
Stock Number:
043915-UCD
Citation ID:
RRID:MMRRC_043915-UCD
Other Names:
hs122/hs123 double enhancer knockout, hs122/hs123 double knockout (KO), hs122/hs123 null, Arx enhancer deletion, ultraconserved enhancer deletion, FVB-Rr43em1AxviRr44em2Axvi

Strain Information

Rr43em1Axvi
Name: regulatory region 43; endonuclease-mediated mutation 1, Axel Visel
Synonyms: deltahs122
Type: Allele
Species: Mus musculus (mouse)
Chromosome: X
Alteration at locus: CRISPR
Rr44em2Axvi
Name: regulatory region 44; endonuclease-mediated mutation 2, Axel Visel
Synonyms: hs122/hs123
Type: Allele
Species: Mus musculus (mouse)
Chromosome: X
Alteration at locus: CRISPR
Rr43
Name: regulatory region 43
Synonyms: hs122
Type: Regulatory Region
Species: Mus musculus (mouse)
Chromosome: X
Alteration at locus: CRISPR
Rr44
Name: regulatory region 44
Synonyms: hs123
Type: Regulatory Region
Species: Mus musculus (mouse)
Chromosome: X
Alteration at locus: CRISPR
Genetic Alterations
This double-KO line was made by injecting single-cell embryos from the hs122 knockout founder lines with the hs123 targeting components. Using CRISPR/Cas9 genome editing, the donor introduced deletions for both the hs122 enhancer (chrX:93,300,959-93,302,702, Rr43) with insertion of "TC" to generate an AatII restriction site in place of the hs122 enhancer) as well as the hs123 enhancer (chrX:92,790,231-92,791,387, Rr44) on the same haplotype. Both enhancers control expression for Arx (MGI:1097716).
Genotype Determination
Phenotype

Homozygous: Homozygous hs122/hs123 females are viable.

Hetero/Hemizygous: Both hs122/hs123 heterozygous-null females and hemizygous-null males are viable and fertile. hs122/hs123 hemizygous-null males display decreased Arx expression in the dorsal forebrain. They also have an abnormal hippocampus, including a dentate gyrus that is substantially reduced in size and has a disorganized appearance.

MeSH Terms
  • Animals
  • Brain/abnormalities
  • Brain/embryology
  • Brain/metabolism
  • Conserved Sequence
  • Embryonic Development/genetics
  • Enhancer Elements, Genetic
  • Female
  • Gene Deletion
  • Homeodomain Proteins/genetics
  • Homeodomain Proteins/metabolism
  • Male
  • Mice
  • Transcription Factors/genetics
  • Transcription Factors/metabolism
Strain Development
The hs122/hs123 double knockout was generated by injecting single-cell embryos from the hs122 (allele Rr43) knockout founder lines with the hs123 (Rr44) targeting components. Using CRISPR/Cas9 genome editing, we introduced deletions for both the hs122 enhancer (chrX:93,300,959-93,302,702) with insertion of "TC" to generate an AatII restriction site in place of the hs122 enhancer) as well as the hs123 enhancer (chrX:92,790,231-92,791,387) on the same haplotype. The hs122 allele was generated by homology-driven repair using a single-stranded oligo template whereas the hs123 allele was generated by nonhomologous end-joining repair. Founders harboring the double-deletions were mated to FVB/NCrl (Charles River Strain 207) for colony expansion and maintenance.
Suggested Control Mice
Wild-type littermates
MMRRC Genetic QC Summary
The MMRRC Centers have developed a genetic QC pipeline using MiniMUGA array genotyping to provide additional information on strain backgrounds for MMRRC congenic and inbred strains. For more information on when data may be available, or to request genotyping for a strain of interest, please contact mmrrc@ucdavis.edu. Older strains may not have this information.
  • Developmental Biology
  • Models for Human Disease
  • Neurobiology
Donor
Axel Visel, Ph.D., Lawrence Berkeley National Laboratory.
Primary Reference

Dickel DE, Ypsilanti AR, Pla R, Zhu Y, Barozzi I, Mannion BJ, Khin YS,Fukuda-Yuzawa Y, Plajzer-Frick I, Pickle CS, Lee EA, Harrington AN, Pham QT,Garvin TH, Kato M, Osterwalder M, Akiyama JA, Afzal V, Rubenstein JLR, PennacchioLA, Visel A. Ultraconserved Enhancers Are Required for Normal Development. Cell. 2018 Jan 25;172(3):491-499.e15. doi: 10.1016/j.cell.2017.12.017. Epub 2018 Jan18. (Medline PMID: 29358049)

Colony and Husbandry Information

Colony Surveillance Program and Current Health 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 mmrrc@ucdavis.edu.
The MMRRC at UC Davis performed microbita analysis on feces samples received from the donating investigator while the mice were still at their facility. There is a MMRRC Fecal Microbiota report available for this strain.If you are interested in having the MMRRC at UC Davis perform microbiota analysis once the line is established at your facility please contact mmrrc@ucdavis.edu.
Coat Color
White (albino)
MMRRC Breeding System
Other or uncertain
Breeding Scheme(s)
Outcrossed to wildtype FVB/NCrl mice
Overall Breeding Performance
Good
NOTE: "Hemizygote" as used here refers to males carrying a mutation on the X Chromosome or mice of either sex carrying an inserted transgene with no homologous allele on the other chromosome.
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes X-linked
Homozygotes are fertile: Undetermined X-linked
Hetero/Hemizygotes are fertile: Yes Yes
Age Reproductive Decline: Undetermined Undetermined
Bred to Homozygosity
No
Average litter size
5-9
Recommended wean age
3 weeks
Average Pups Weaned
5-9

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 mmrrc@ucdavis.edu 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
043915-UCD-EMBRYO Cryo-preserved embryos $1,038.00 / Non-Profit Aliquot Approximate quantity2 : 20-40 embryos / aliquot
043915-UCD-SPERM Cryo-preserved spermatozoa $546.25 / Non-Profit Aliquot Approximate quantity3
043915-UCD-RESUS Litter recovered from cryo-archive $4,044.00 / Non-Profit Litter Recovered litter4; additional fees for any special requests.

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.