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Genomic DNA including Car3 was isolated from a mouse 129SvJ genomic library, and the genomic organization of Car3 was determined by restriction digestions and sequencing. The Car3 gene targeting construct was generated by subcloning a 4.0-kb NotI/XbaI genomic fragment containing the 5’ end of the locus and a 6.7-kb SpeI/NheI fragment containing the 3’ end in a pPNT backbone. The resulting vector replaced the 4.2-kb XbaI/SpeI fragment containing exons 3, 4, and 5 of Car3 with a PGKneo gene. The targeting vector was linearized with NotI and transfected into TC1 embryonic stem cells (derived from 129S6/SvEvTac) by electroporation. One hundred eighteen colonies resistant to geneticin were analyzed for homologous recombination by Southern blotting and PCR. Eight clones were selected and verified by PCR and extensive restriction analysis with the use of flanking probes to demonstrate that the locus was targeted correctly. Two clones were injected into C57BL/6J blastocysts and implanted into (CBA x C57BL/6J)F1 foster mothers. Male chimeras from two clones selected by agouti color were mated to C57BL/6 females to confirm germline transmission. Agouti offspring were tested for the targeted Car3 allele by Southern blotting or PCR of tail DNA. Chimeric males were bred to wild-type 129SvEv females to generate mice heterozygous for the Car3 deletion in a pure 129SvEv background.
(Adopted from PMID:15509796)
Kim G, Lee TH, Wetzel P, Geers C, Robinson MA, Myers TG, Owens JW, Wehr NB,Eckhaus MW, Gros G, Wynshaw-Boris A, Levine RL. Carbonic anhydrase III is notrequired in the mouse for normal growth, development, and life span. Mol CellBiol. 2004 Nov;24(22):9942-7. (Medline PMID: 15509796)
Colony Surveillance Program and Current Health 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 mmrrc@med.unc.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.
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.