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RetnlaTdT constitutive, RELMalpha knockout/tDTomato reporter knock-in (constitutive)
Cre-Lox/FLEx technology replaced RELMalpha (Retnla) with a tdTomato reporter. More specifically, the reporter cassette was inserted in antisense orientation downstream of exon 4. Upon Cre-recombination, the reporter gene was inverted, into sense orientation, which simultaneously deleted exon Renlna exons 2-4. A woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) enhances reporter expression (PMID:12787924).
Mice are viable, fertile, and overall seem healthy under normal vivarium conditions. However, they have more infection-induced inflammation in the intestine and serosal cavity, and more fibrosis and granulomas.
Commercial transgenic mouse services (genOway) combined Cre-Lox and FLEx technology to delete the RELMalpha gene (Retnla) and replace it with a functional tdTomato reporter on a C57BL/6 background. More specifically: electroporation of the targeting construct into ES cells (proprietary of genOway derived from C57BL/6N), neomycin selection, clonal expansion, and screening via Southern for correct targeting and Flp/Cre-mediated excision of Retnla locus and insertion of the IRES tdTomato-WPRE cassette. Six ES clones injected into blastocysts; implantation into OF1 females to produce chimeras. Tail-snip PCR to confirm the chimera genotype and mating of offspring with C57BL/6N (Charles River) for germline transmission. Three founders were identified and bred with C57BL/6J (The Jackson Laboratories) mice for two generations. The line was then crossed to a tissue-specific Cre-epxressing strain (B6.Cg-Tg(Itgax-cre)1-1Reiz/J, RRID:IMSR_JAX:008068). This conditional KO line was crossed to a proprietary, ubiquitous Cre-deleter line (genOway, CMV-driven Cre) to create a constitutive RELMalpha knockout with the tdTomato knock-in.
Note: This line is available, conditional-ready, in MMRRC:67029.
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@ucdavis.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.
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.