Description

Strain Information

Type Coisogenic; Mutant Strain; Targeted Mutation; Additional information on Genetically Engineered Mutant Mice. Mating System Homozygote x Homozygote         (Female x Male) Species laboratory mouse Generation N21+ (21-JUL-08)   Donating Investigator Philip Ashton-Rickardt,   Imperial College London

Description
Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. No gene product (mRNA) is detected by RT-PCR analysis of myeloid and lymphoid cells. After experimental viral infection, homozygous mice have lower (five to nine-fold reduction) numbers of virus specific cytotoxic T lymphocytes (CTLs) and increased apoptosis of virus specific CD8 T cells when compared to wildtype controls. Mutant mice have impaired Lymphocytic Choriomeningitis virus (LCMV) infection clearance. Granzyme A and granzyme B specific activity is reduced in cytotoxic granules and increased in CTL cytoplasm. Mutant CTLs have a three-fold increase in the number of granules lacking granzyme B. This mutant mouse strain may be useful in studies of viral immune response and inflammation, and the exocytosis pathway in CTL apoptosis.

Development
A targeting vector containing a floxed neomycin resistance selection cassette was used to disrupt exon 7, which contains most of the coding sequence. The construct was electroporated into C57BL/6-derived embryonic stem (ES) cells (Ware et al. Transgenic Res. 12, 743.), which were transiently transfected with a Cre recombinase vector to remove the selection cassette. ES cells that had successfully undergone Cre-mediated recombination and no longer retained the cassette were injected into BALB/c blastocysts. The resulting chimeric animals were crossed to C57BL/6J mice, and then backcrossed to the same for 20 generations.

Control Information

	Control
	000664 C57BL/6J
Considerations for Choosing Controls

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms

      assigned by genotype

Serpinb9^tm1Arp/Serpinb9^tm1Arp

        involves: C57BL/6

• cellular phenotype
• increased apoptosis (MGI Ref ID J:113349)
  • apoptosis is increased in LCMV- and LM-specific CTLs in mutants
• hematopoietic system phenotype
• abnormal CD8-positive, alpha-beta cytotoxic T cell morphology (MGI Ref ID J:113349)
  • upon infection with LCMV Armstrong or a strain of Listeria monocytogenes (LM), numbers of LCMV-specific CTLs are reduced 5-fold and LM-specific CTLs are reduced 9-fold compared to control B6 mice
  • frequency of CTLs devoid of Gsmb granules is 3 times greater than in controls; number of Gzmb granules per CTL is 2-fold lower than in controls
• homeostasis/metabolism phenotype
• abnormal enzyme/ coenzyme level (MGI Ref ID J:113349)
  • caspase 3 in cytoplasm is increased
• abnormal enzyme/coenzyme activity (MGI Ref ID J:113349)
  • GranzymeB activity is significantly increased in cytoplasm of mutants compared to controls
• immune system phenotype
• abnormal CD8-positive, alpha-beta cytotoxic T cell morphology (MGI Ref ID J:113349)
  • upon infection with LCMV Armstrong or a strain of Listeria monocytogenes (LM), numbers of LCMV-specific CTLs are reduced 5-fold and LM-specific CTLs are reduced 9-fold compared to control B6 mice
  • frequency of CTLs devoid of Gsmb granules is 3 times greater than in controls; number of Gzmb granules per CTL is 2-fold lower than in controls
• abnormal cytotoxic T cell physiology (MGI Ref ID J:113349)
  • apoptosis is increased in LCMV- and LM-specific CTLs in mutants
• defective cytotoxic T cell cytolysis (MGI Ref ID J:113349)
  • deficit in CTL activity is observed in ex vivo activity assay over course of LCMV infection; on day 10, activity is decrease 6-fold compared to controls
• increased susceptibility to viral infection (MGI Ref ID J:113349)
  • clearance of LCMV is defective in mutants; titer of virus is 10-fold higher on day 6 of infection compared to wild-type

View Research Applications

Research Applications

This mouse can be used to support research in many areas including:

Apoptosis Research
Extracellular Modulators

Immunology and Inflammation Research
Growth Factors/Receptors/Cytokines

Research Tools
Apoptosis Research

Genes & Alleles

Gene & Allele Information

Allele Symbol		Serpinb9tm1Arp
Allele Name		targeted mutation 1, Philip G Ashton-Rickardt
Allele Type		Targeted (knock-out)
Common Name(s)		M Spi6 delta Neo; Spi6-;
Mutation Made By		Philip Ashton-Rickardt,   Imperial College London
Strain of Origin		C57BL/6
Gene Symbol and Name		Serpinb9, serine (or cysteine) peptidase inhibitor, clade B, member 9
Chromosome		13
Gene Common Name(s)		BB283241; CAP-3; CAP3; MGC94010; PI-9; PI9; Spi6; expressed sequence BB283241; ovalbumin; serine protease inhibitor 6;
Molecular Note		Exon 7 was replaced with a floxed neomycin resistance cassette that was subsequently removed. [MGI Ref ID J:113349]

Genotyping

Genotyping Information

Genotyping Protocols

Serpinb9^tm1Arp, SEP PCR, vers. 1

Helpful Links

Optimizing PCR Protocols

References

References

Selected Reference(s)

Zhang M; Park SM; Wang Y; Shah R; Liu N; Murmann AE; Wang CR; Peter ME; Ashton-Rickardt PG. 2006. Serine protease inhibitor 6 protects cytotoxic T cells from self-inflicted injury by ensuring the integrity of cytotoxic granules. Immunity 24(4):451-61. [PubMed: 16618603]  [MGI Ref ID J:113349]

Health & husbandry

Health & Colony Maintenance Information

Animal Health Reports

Room Number           AX11

Colony Maintenance

Breeding & Husbandry	When maintaining a live colony, these mice can be bred as homozygotes.
Mating System	Homozygote x Homozygote         (Female x Male)
Diet Information	LabDiet® 5K52/5K67

Purchasing information

Pricing, Supply Level & Notes, Controls, General Terms & Conditions

Pricing

Supply Details

Standard Supply

Repository-Live. A collection of over 1000 strains maintained as live colonies. Individual colonies are sized to meet current customer demand. Delivery for orders of 10 mice or less ranges on average from one to eight weeks; mice are generally shipped between four to six weeks of age with a maximum shipping age of ~nine weeks. Colony sizes do not generally support stringent age specifications for large volumes of mice; however custom orders and larger quantities of mice are easily arranged. Estimated ship dates for all orders provided within 48 hours of order placement.

Supply Notes

Usually shipped between four and eight weeks of age. This strain is included in the Induced Mutant Resource Colony collection.

Control Information

	Control
	000664 C57BL/6J
Considerations for Choosing Controls
USA, Canada and Mexico - Control Pricing Information for Genetically Engineered Mutant Strains.
International - Control Pricing Information for Genetically Engineered Mutant Strains.

General Terms and Conditions

See Terms of Use

The Jackson Laboratory's Genotype Promise

The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX^® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX^® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX^® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project.

---

Ordering and Purchasing Information

      Purchasing Information
      JAX^® Mice Orders
      Surgical Services

Contact Information
Orders & Technical Support
Tel: 800.422.6423 or 207.288.5845
Fax: 207.288.6150
Technical Support Email Form

Terms of Use

Terms of Use

General Terms and Conditions

Effective September 26, 2007: License Requirements for Strains using Cre-lox Technology only apply in Canada, see Licenses for Strains using Cre-lox Technology.

Contact information

General inquiries

Contracts Administration

phone:	207-288-6470
fax:	207-288-6655

JAX^® Mice & Services Conditions of Use

“Each recipient institution, including its employees and other researchers under its control (RECIPIENT), of mice or services using mice from The Jackson Laboratory (TJL) agrees that such mice, descendants of those mice derived by inbreeding or crossbreeding, including unmodified derivatives of those mice or their descendants (“MICE”) shall not be: (i) used for any purpose other than the internal research of the RECIPIENT, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services with respect to MICE. Acceptance of MICE from TJL shall be deemed agreement by RECIPIENT to these conditions, and departure from these conditions requires The Jackson Laboratory’s prior written authorization.”

In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of MICE, products or services, The Jackson Laboratory will, at its option, provide credit or replacement for the MICE or product received or the services provided.

No Liability

In no event shall The Jackson Laboratory, its trustees, directors, officers, employees, and affiliates be liable for any causes of action or damages, including any direct, indirect, special, or consequential damages, arising out of the provision of MICE, products or services, including economic damage or injury to property and lost profits, and including any damage arising from acts or negligence on the part of The Jackson Laboratory, its agents or employees. In purchasing or receiving MICE, products or services from The Jackson Laboratory, purchaser or recipient, or any party claiming by or through them, expressly releases and discharges The Jackson Laboratory from all such causes of action or damages, and further agrees to defend and indemnify The Jackson Laboratory from any costs or damages arising out of any third party claims.

MICE and biological materials are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.

The foregoing represents the General Terms and Conditions applicable to The Jackson Laboratory’s MICE, products and services. In addition, special terms and conditions of sale of certain MICE, products and services may be set forth separately in The Jackson Laboratory web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, products and services by The Jackson Laboratory, and by its licensees and distributors.

Acceptance of delivery of MICE, products or services shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on The Jackson Laboratory, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, products services by The Jackson Laboratory.