Strain Name:
C57BL/6J-Gadd45aem1Ahoke/Mmjax
Stock Number:
050634-JAX
Citation ID:
RRID:MMRRC_050634-JAX
Other Names:
C57BL/6J-Gadd45tm1Ahoke

Strain Information

Gadd45aem1Ahoke
Name: growth arrest and DNA-damage-inducible 45 alpha; endonuclease-mediated mutation 1, Ahmet Hoke
Synonyms: Gadd45aflox
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 6
Alteration at locus: CRISPR
Gadd45a
Name: growth arrest and DNA-damage-inducible 45 alpha
Synonyms: Ddit1
Type: Gene
Species: Mouse
Chromosome: 6
Alteration at locus: CRISPR
NCBI: 13197
HGNC: HGNC:4095
Homologene: 1449
Genetic Alterations
CRISPR/Cas9 endonuclease-mediated genome editing was used to insert loxP sites flanking the first three coding exons of the gene. In addition, a V5 epitope tag was placed directly after the start codon.
Genotype Determination
Phenotype
None/Normal/Wild-type
Strain Development
  • Guide RNAs targeting intronic regions flanking the first three coding exons of Gadd45a were designed using the web-based CRISPR design tool of MIT (crispr.mit.edu). Two of the top gRNAs (score ≥ 90) were selected for use as follows: Gadd45a-gRNA-5’: 5’-CAGCACCACCTTCGTCC^CGT -3’; Gadd45a-gRNA-3’: 5’-CGGAGCGTGTCTAAGCT^CGT -3.
  • Alt-R CRISPR-Cas9 crRNA oligos and a 1.3 kb single-stranded donor DNA megamer containing the desired 5’ loxP, 3’ loxP, and a V5 epitope tag introduced directly after the start codon were synthesized by IDT (Coralville, IA).
  • Pronuclear injection of one-cell C57BL/6J embryos (Jackson Laboratories) was performed by the JHU Transgenic Core using standard microinjection techniques using a mix of Cas9 protein (30 ng/ul, PNABio), tracrRNA (0.6 µM, Dharmacon), crRNA (0.6 µM, IDT) and ssDNA oligo (5 ng/ul, IDT) diluted in RNAse-free injection buffer (10 mM Tris-HCl, pH 7.4, 0.25 mM EDTA).
  • Injected embryos were transferred into the oviducts of pseudopregnant ICR females (Envigo).
  • 5’ loxP and V5 epitope insertion in resulting pups was validated using the following primers: Gadd45a-loxP-F3: 5’-AGGACACTTGAACCACTGCAA-3’; Gadd45a-V5-R: 5’-CAGGCACACTTACCTTTCGGT-3’; expected product sizes 510 bp (unmodified) and 586 bp (modified). 3’ loxP insertion was validated using the following primers: Gadd45a-loxP-F1: 5’-TGCTGCTACTGGAGAACGAC-3’; Gadd45a-loxP-R2: 5’-AATTAGCCACGCGAGGTTGT-3’; expected product sizes 285 bp (unmodified) and 319 bp (modified).
  • PCR fragments from candidate founding lines were purified using AMPure XP beads (Beckman Coulter) and verified by Sanger sequencing.
  • A homozygous founder carrying both modified sites is designated C57BL/6J-Gadd45atm1Ahoke and was backcrossed 3 generations to C57BL/6J mice, then submitted to the MMRRC.
Suggested Control Mice
C57BL/6J
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 csmmrrc@jax.org. Older strains may not have this information.
  • Apoptosis
  • Cancer
  • Cell Biology
  • Developmental Biology
  • Immunology and Inflammation
  • Neurobiology
  • Research Tools
Donor
Ahmet Hoke, M.D., Johns Hopkins School of Medicine.

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
N3 (C57BL/6J)
Overall Breeding Performance
Good
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Yes Yes
Heterozygotes are fertile: N/A N/A
Age Reproductive Decline: Undetermined Undetermined
Bred to Homozygosity
Yes
Average litter size
4-6
Recommended wean age
3 Weeks
Average Pups Weaned
4-6

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
050634-JAX-SPERM Cryo-preserved spermatozoa $437.00 / Non-Profit Aliquot Approximate quantity3
050634-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.