Strain Name:
C57BL/6-Kif9em1.1Seje/Mmjax
Stock Number:
066701-JAX
Citation ID:
RRID:MMRRC_066701-JAX
Other Names:
Kif9 knockout

Strain Information

Kif9em1.1Seje
Name: kinesin family member 9; endonuclease-mediated mutation 1.1, Jeffrey Segall
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 9
Alteration at locus: CRISPR
Kif9
Name: kinesin family member 9
Type: Gene
Species: Mouse
Chromosome: 9
Alteration at locus: CRISPR
NCBI: 16578
Homologene: 65620
Genetic Alterations
Targeted deletion of exon 3
Genotype Determination
ES Cell Line
Not applicable
Phenotype
Male sterility in homozygous knockout animals. The homozygous floxed (MMRRC:66522) also has a slightly higher litter size compared to a male het KO x female homozygous KO.
Strain Development
A pair of gRNAs targeting intron 2 (gRNAE22) and intron 3 (gRNAE23) of Kif9 gene respectively were designed using an online tool (https://benchling.com/) and generated by in vitro transcription. Cas9 protein was purchased from PNA Bio Inc. A Kif9 conditional knockout HRD plasmid carrying around 2-kb homologous arms on each side surrounding a floxed Kif9 exon3 with an added primer sequence for genotyping was generated by SLiCE cloning. Superovulated female C57BL/6 mice (3–4 weeks old) were mated to C57BL/6 males, and fertilized embryos were collected from oviducts. The gRNAs targeting introns 2 and intron 3 of the Kif9 gene, the Cas9 protein, and the Kif9 conditional knockout HRD were mixed and microinjected into the pronuclei of fertilized eggs. The injected zygotes were transferred into pseudopregnant CD1 females, and the resulting pups were genotyped. Founder mice carrying floxed Kif9 alleles were identified by PCR using primers 3’SR and 5’SF and sequencing. Founders were mated with C57BL/6 mice to get F1 offspring. Crossing of F1 floxed heterozygotes was used to generate homozygous floxed mice. Knockout mice were generated by breeding floxed homozygotes with the Cre deleter strain C57BL/6-Tg(Zp3-cre)93Knw/J (RRID:IMSR_JAX:003651). Genotyping was performed by PCR of genomic DNA using the 3’SR and 5’SF primers, with product sizes used to determine genotype: 1003 bp for WT, 1085 bp for floxed; the knockout should produce a 593-bp band.
Suggested Control Mice
C57BL/6J
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 csmmrrc@jax.org. Older strains may not have this information.
  • Cell Biology
  • Reproduction
Donor
Jeffrey Segall, Ph.D., Albert Einstein College of Medicine.
Primary Reference

Chen K, Cho SY, Zhang Y, Beck A, and Segall JE, Bioarchives - https://doi.org/10.1101/2020.03.21.001602

Colony and Husbandry Information

Cryo-recovered strains distributed by the MMRRC at JAX are shipped to the customer from the Pathogen & Opportunistic-Free Animal Room G200 - see https://www.jax.org/jax-mice-and-services/customer-support/customer-service/animal-health/health-status-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 csmmrrc@jax.org.
Coat Color
Black
Eye
Black
MMRRC Breeding System
Sib-mating
Generation
N/A
Overall Breeding Performance
Excellent
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Yes Sterile
Heterozygotes are fertile: Yes Yes
Age Reproductive Decline: Undetermined Undetermined
Average litter size
4 to 6
Recommended wean age
4 Weeks
Average Pups Weaned
4 to 6

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 csmmrrc@jax.org 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.

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
066701-JAX-SPERM Cryo-preserved spermatozoa $437.00 / Non-Profit Aliquot Approximate quantity3
066701-JAX-RESUS Litter recovered from cryo-archive $2,022.00 / Non-Profit Litter Recovered litter4; additional fees for any special requests.
Cryopreserved material may be available upon request, please inquire to csmmrrc@jax.org 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.