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129P2/OlaHsd-derived E14 ES cells were transfected by electroporation and G418-resistant clones were isolated. Correctly targeted clones were identified by Southern hybridization to purified genomic DNA using an upstream probe following digestion by NdeI and XhoI and using a downstream probe following digestion by XbaI. A correctly targeted ES cell subclone (1A10) was used for blastocyst injection and embryo transfer. One high chimera male was obtained and mated to a C57BL/6J female to generate heterozygous targeted mice (y-addCKO/+), initially identified by neor gene PCR, and confirmed by 5’ and 3’ Southern analysis of tail DNA, as described above. The neor gene was excised by mating a correctly targeted heterozygous mouse to 129S4/SvJaeSor-Gt(ROSA)26Sortm1(FLP1)Dym/J; (JAX Strain #003946); y-addneo/+ pups were identified by PCR. The floxed exon 2 was excised by mating a y-addneo/+ mouse to B6.FVB-Tg(EIIa-cre)C5379Lmgd/J; (JAX Strain #003724), producing heterozygous mice with the targeted y-adducin allele lacking both exon 2 and the neor cassette.
Sahr KE, Lambert AJ, Ciciotte SL, Mohandas N, Peters LL. Targeted deletion of the gamma-adducin gene (Add3) in mice reveals differences in alpha-adducininteractions in erythroid and nonerythroid cells. Am J Hematol. 2009Jun;84(6):354-61. doi: 10.1002/ajh.21427. PubMed PMID: 19425068; PubMed Central(Medline PMCID: PMC2827150)
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.
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.
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.