Register New MMRRC Account
Reset Password
Register for New Mouse Models
Click Here for Additional Contact Information
Availability & Fees Order this Strain
Homozygous: Normal.
Hetero/Hemizygous: Normal.
Clones spanning the Ptpn11 locus were isolated from a 129/SvJ genomic library. A targeting vector was constructed by inserting loxP sites to flank Ptpn11, exon11 (Fig. S1), followed by a splice acceptor b-galactosidase/pGK-neo cassette (SAbgal pGK/Neo). The diphtheria toxin-A gene also was introduced to facilitate negative selection.
Mouse ESVJ-1182 ES cells (Go Germline; Genome Systems) were cultured on irradiated mouse embryonic fibroblasts (MEFs) in ES cell medium (Dulbecco’s modified eagle's medium containing leukemia inhibitory factor; Gibco BRL, Rockville, MD) at 37°C and 5% CO2. ES cell propagation, electroporation and selection of recombinants in G418-containing media, and preparation and culture of MEFs and bone marrow-derived macrophages (BMMs) were carried out as described previously (PMID:16516835 and PMID:8524272). Three correctly targeted ES clones (#49, #79, and #183) were first identified by PCR screening and then confirmed by Southern blots with probe A after digestion with SpeI (b, left). Clones #49 and #79 were later electroporated with the plasmid pGK-Cre. Upon screening, two recombinant clones (J40, I22) carrying the Ptpn11 floxed allele were identified by PCR and confirmed by Southern blot analysis of EcoR1-digested genomic DNA, using probes B and C (Fig. S1). Chimeras were generated by injecting two of the recombinant ES cell clones into C57BL/6J blastocysts. High percentage chimeras from both clones (which behaved identically) were crossed to C57BL/6J mice to generate heterozygotes. Mice were genotyped for the floxed (430 bp) and wild-type Ptpn11 (312 bp) alleles by PCR (Forward primer F1: 5’-TAG CTG CTT TAA CCC TCT GTG T; Reverse primer R1: 5’-CAT CAG AGCAGG CCA TAT TCC) using tail DNA.
Mice carrying one Ptpn11-deleted allele (ΔK11) were generated by germline deletion of the floxed Ptpn11 allele, following crossing to actin-Cre mice (PMID:9598348). MEFs bearing one Ptpn11Δk11 allele were derived from E13.5 embryos, as described earlier (PMID:16516835).
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.
The donor or their institution limits the distribution to non-profit institutions only.
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.
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.