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Six6flox mice were generated from a Six6 European Conditional Mouse Mutagenesis Program(EUCOMM) KO first construct. Because the Six6 gene has only two exons and thus only one intron, an artificial intron was inserted into exon 1 to contain the flippase recombinase target(FRT)–flanked selection cassette followed by the 5' loxPsite. The intron was derived from the Ifitm2 gene and remains in the final Six6 exon 1 in the floxed mouse, where it contains the remaining single FRT element and the 5' loxP site. The plasmid was introduced by electroporation into male C57BL/6J mouse embryonic stem cells, selected by using G418 (selection against vector insertion was diphtheria toxin A on the backbone of the plasmid), and screened for homologously recombined cells by PCR. A homologously recombined clone of mouse embryonic C57BL/6J stem cells was then injected into albino C57BL/6J blastocysts to produce chimeric mice. These were screened for percentage chimerism and correct recombination. The highest percentage male chimeric mice were bred to C57BL/6J females to create heterozygous Six6flox mice. Once confirmed as correctly floxed for Six6, they were crossed with a FLPase mouse (B6;SJL-Tg(ACTFLPe)9205Dym/J - RRID:IMSR_JAX:003800) to remove the selection cassette, leaving only one FRT site and the 5' loxP site in the artificial intron and the 3' loxP site in the original intron between exons 1 and 2 to create the Six6flox conditional KO allele. LoxP sites flanked most of exon 1 of the Six6 gene. Floxed homozygous mice evidenced no previously described phenotypes, indicating that the artificial intron was correctly spliced.
Conditional phenotype: Yes, various when crossed with specific Cre alleles.
Pandolfi EC, Tonsfeldt KJ, Hoffmann HM, Mellon PL. Deletion of the Homeodomain Protein Six6 From GnRH Neurons Decreases GnRH Gene Expression, Resulting in Infertility. Endocrinology. 2019 Sep 1;160(9):2151-2164. doi: 10.1210/en.2019-00113. (Medline PMID: 31211355)
Pandolfi EC, Breuer JA, Nguyen Huu VA, Talluri T, Nguyen D, Lee JS, Hu R, Bharti K, Skowronska-Krawczyk D, Gorman MR, Mellon PL, Hoffmann HM. The Homeodomain Transcription Factors Vax1 and Six6 Are Required for SCN Development and Function. Mol Neurobiol. 2020 Feb;57(2):1217-1232. doi: 10.1007/s12035-019-01781-9. Epub 2019 Nov 9. (Medline PMID: 31705443)
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.
A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Devora Rossi.
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 Devora Rossi
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.