Strain Name:
B6N.B6J(SJL)-Six6tm1.1Plm/PmelMmucd
Stock Number:
068240-UCD
Citation ID:
RRID:MMRRC_068240-UCD
Other Names:
Six6 Flox

Strain Information

Six6tm1.1Plm
Name: sine oculis-related homeobox 6; targeted mutation 1.1, Pamela L Mellon
Synonyms: Six6flox
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 12
Alteration at locus: Conditional
Six6
Name: sine oculis-related homeobox 6
Synonyms: Six9, Optx2
Type: Gene
Species: Mouse
Chromosome: 12
Alteration at locus: Conditional
NCBI: 20476
Homologene: 7220
Genetic Alterations

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.

ES Cell Line
Not specified
Phenotype
No phenotype unless crossed with a Cre allele.

Conditional phenotype: Yes, various when crossed with specific Cre alleles.

MeSH Terms
  • Animals
  • Apoptosis
  • Gonadotropin-Releasing Hormone/genetics
  • Homeodomain Proteins/physiology
  • Hypogonadism/etiology
  • Infertility/etiology
  • Male
  • Mice, Inbred C57BL
  • Trans-Activators/physiology
  • Circadian Rhythm/physiology
  • Gene Expression Regulation/physiology
  • Gonadotropin-Releasing Hormone/metabolism
  • Homeodomain Proteins/metabolism
  • Hypothalamus/metabolism
  • Mice
  • Neurons/metabolism
  • Neuropeptides/metabolism
  • Suprachiasmatic Nucleus/growth & development
  • Suprachiasmatic Nucleus/physiology
  • Trans-Activators/metabolism
  • Transcription Factors/metabolism
Strain Development
See details in the "Genetic Alterations" section. The mice were backcrossed for many years to C57BL/6NHsd.
Suggested Control Mice
C57BL/6N
MMRRC Genetic QC Summary
The MMRRC Centers have developed a genetic QC pipeline using MiniMUGA array genotyping to provide additional information on strain backgrounds for MMRRC congenic and inbred strains. For more information on when data may be available, or to request genotyping for a strain of interest, please contact mmrrc@ucdavis.edu. Older strains may not have this information.
  • Developmental Biology
  • Neurobiology
  • Reproduction
Donor
Pamela Mellon, Ph.D., University of California, San Diego.
Primary Reference

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 and Husbandry Information

Colony Surveillance Program and Current Health Reports

Mice recovered from a cryo-archive will have health surveillance performed on recipient females. Health reports will be provided prior to shipment. If you require additional health status information, please email mmrrc@ucdavis.edu.
Coat Color
Black
Eye
Black
MMRRC Breeding System
Backcross
Generation
N5+ (C57BL/6NHsd)
Overall Breeding Performance
Excellent
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Yes Yes
Heterozygotes are fertile: Yes Yes
Age Reproductive Decline: 7 to 9 months 10 to 12 months
Average litter size
7 to 9
Recommended wean age
3 Weeks
Average Pups Weaned
7 to 9

Order Request Information

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.

Click button to Request this one strain. (Use the MMRRC Catalog Search to request more than one strain.)
MMRRC Item # Description Distribution Fee / Unit (US $)
*Shipping & Handling not included*
Units Notes
068240-UCD-SPERM Cryo-preserved spermatozoa $546.25 / $869.88
Non-Profit / For-Profit
Aliquot Approximate quantity3
068240-UCD-RESUS Litter recovered from cryo-archive $4,044.00 / $7,650.23
Non-Profit / For-Profit
Litter Recovered litter4; additional fees for any special requests.
Cryopreserved material may be available upon request, please inquire to mmrrc@ucdavis.edu for more information.

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.

To request material from the MMRRC: Please fill out our on-line request form (accessible from the catalog search results page, or click the Request this Strain button in the fees section). If you have questions or need assistance completing this form, you may call Customer Service at (800) 910-2291 (in USA or Canada) or (530) 757-5710 (international calls). Before you call, please have with you: the MMRRC item number, quantity needed, Bill-to and Ship-to contact information.