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A 2.2-kb targeting fragment spanning exons 6∼9 of mouse Fam20c was produced to create the Fam20c-floxed allele. The introduction of Cre recombinase will excise exons 6-9. The neo cassette was removed by transient transfection with an Flp-expressing vector.
To generate the Fam20c-floxed allele, a 2.2-kb targeting fragment spanning exons 6∼9 of mouse Fam20c was PCRed from genomic DNA of WW6 ES cells. The fragment was subcloned upstream of an Frt-flanked mcl-Neo cassette in the conditional targeting vector pFlox-Frt-Neo. Homologous arms were generated by PCR and subcloned into the targeting vector along with a downstream negative selection PGK-DTA (diphtheria toxin A) cassette. The final targeting construct was electroporated into W4 ES cells (129S6/SvEvTac) and clones were picked after G418 selection. Appropriately targeted (PCR) ES cell clones transiently transfected with the pCAGGS-flpE-puro vector (RRID:Addgene_20733) to remove the neo cassette, which was confirmed by PCR. No random integration was noted. The cells were injected into C57BL/6 blastocysts and male chimeras were crossbred with C57BL/6J females to produce F1 agouti offspring. Genotyping was done via PCR to breed heterozygotes to homozygosity. The resulting Fam20cflox/flox breeding pairs were maintained hom/hom breeding.
Wang X, Wang S, Li C, Gao T, Liu Y, Rangiani A, Sun Y, Hao J, George A, Lu Y, Groppe J, Yuan B, Feng JQ, Qin C. Inactivation of a novel FGF23 regulator,FAM20C, leads to hypophosphatemic rickets in mice. PLoS Genet.2012;8(5):e1002708. doi: 10.1371/journal.pgen.1002708. Epub 2012 May 17. PubMedPMID: 22615579; PubMed Central (Medline PMCID: PMC3355082)
Colony Surveillance Program and Current Health Reports
Limited quantities of breeder mice (up to 2 males and 2 females or 4 mice) per investigator per month are available from a live colony, usually available to ship in under 12 weeks. Larger quantities may be available, please contact the distributing center directly at mmrrc@missouri.edu for more details.
Cryopreserved material may be available upon request, please inquire to mmrrc@missouri.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.