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A targeting vector was designed containing a frt-flanked neomycin resistance (neo) cassette followed by loxP sites flanking Muc5a exon 1 and upstream sequence. The construct was electroporated into 129S1-derived CJ7 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts, and the resulting chimeras were bred to C57BL/6J mice. Knockout mice were then produced by mating founder animals with Zp3-Cre transgenic mice (C57BL/6-Tg(Zp3-cre)93Knw/J).
A targeting vector was designed containing a frt-flanked neomycin resistance (neo) cassette followed by loxP sites flanking Muc5a exon 1 and upstream sequence. The construct was electroporated into 129S1-derived CJ7 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts, and the resulting chimeras were bred to C57BL/6J mice. Knockout mice were then produced by mating founder animals with Zp3-Cre transgenic mice (C57BL/6-Tg(Zp3-cre)93Knw/J) and subsequently crossing progeny with C57BL/6J mice. Mice were backcrossed to C57BL/6J for eight generations, and were further crossed to C57BL/6NTac background mice for 12 generations.
Wild type or heterozygous mice from the colony may be used as controls. C57BL/6NTac may also be used.
Roy MG, Livraghi-Butrico A, Fletcher AA, McElwee MM, Evans SE, Boerner RM, Alexander SN, Bellinghausen LK, Song AS, Petrova YM, Tuvim MJ, Adachi R, Romo I, Bordt AS, Bowden MG, Sisson JH, Woodruff PG, Thornton DJ, Rousseau K, De la Garza MM, Moghaddam SJ, Karmouty-Quintana H, Blackburn MR, Drouin SM, Davis CW, Terrell KA, Grubb BR, O'Neal WK, Flores SC, Cota-Gomez A, Lozupone CA, Donnelly JM, Watson AM, Hennessy CE, Keith RC, Yang IV, Barthel L, Henson PM, Janssen WJ, Schwartz DA, Boucher RC, Dickey BF, Evans CM. Muc5b is required for airway defence. Nature. 2014 Jan 16;505(7483):412-6. doi: 10.1038/nature12807. Epub 2013 Dec 8. (Medline PMID: 24317696)
Hasnain SZ, Evans CM, Roy M, Gallagher AL, Kindrachuk KN, Barron L, Dickey BF, Wilson MS, Wynn TA, Grencis RK, Thornton DJ. Muc5ac: a critical component mediating the rejection of enteric nematodes. J Exp Med. 2011 May 9;208(5):893-900. doi: 10.1084/jem.20102057. Epub 2011 Apr 18. (Medline PMID: 21502330)
After these mice were backcrossed 12 generations onto the C57BL6NTac background, the donating investigator maintained the line as a het x het cross to get all of the genotypes or a ko x ko cross to maintain the line.
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.
A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Carmen Melvin.
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.
A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Carmen Melvin
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.
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