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 N10F?+ (27-MAY-08)   Donating Investigator Motomu Shimaoka,   Immune Disease Institute (formerly CBRI)

Description
Mice homozygous for this b₇ (D146A) targeted mutation are viable and fertile. DNA sequencing confirms an aspartate (D) to alanine (A) substitution at position 146 of the targeted b₇ integrin gene. As such, homozygotes have a mutant ADMIDAS (adjacent to metal ion-dependent adhesion site) cation binding/exchange site in the b₇ integrin head (A-domain). The resulting imbalance between the non-adhesive and adhesive states of leukocyte integrins skews toward a persistently adhesive state. Homozygous mutants exhibit impaired leukocyte migration, perturbed lymphocyte trafficking in the gut, and reduced T- and B-cell numbers in small/large bowel and gut-associated lymphoid tissues (less T-cells in Peyer's patches (PP), fewer intraepithelial lymphocyte (IEL) and lamina propria lymphocyte (LPL) compartments in the small intestine; and less B-cells in PP and the large intestine). In addition, CD4⁺CD45RB^high T cells isolated from b₇ (D146A) homozygotes have reduced colitogenic potential compared to CD4⁺CD45RB^high T cells taken from wildtype mice following adoptive transfer. As the mouse integrin b₇ chain is found as a cell surface heterodimer in association with either of two alpha chains (a₄ or a_E), these b₇ (D146A) mutant mice may be useful in studying intestinal leukocyte migration and trafficking, integrin/ligand interactions (VCAM-1, fibronectin, MadCAM-1, E-cadherin), intraepithelial lymphocytes, and inflammatory bowel diseases.

Of note, many strains harboring integrin beta mutant alleles are available from The Jackson Laboratory, including Itgb1-deficient (beta-1 null; Stock No. 003096), Itgb1-flox (Stock No. 004605), Itgb2-hypomorph (CD18-hypo; Stock No. 002128), Igtb2-deficient (CD18-null or beta2-null; Stock No. 003329), Itgb3-deficient (beta3-null; Stock No. 004669), Itgb5-deficient (beta5-null; Stock No. 004166), Itgb7-deficient (beta7-null; Stock No. 002965 and Stock No. 004944), and Itgb7^D146A mutant (Stock No. 007707) strains.

Development
To generate this mutant allele, an ACN targeting vector was designed to insert (by site-directed mutagenesis) a single amino acid substitution of aspartic acid to alanine at codon 146 (D146A) into exon 4, and an engineered EcoRI site downstream of exon 4 of the targeted gene. The ACN cassette, containing the neomycin resistance gene and Cre recombinase gene under the control of the sperm-specific angiotensin-converting enzyme promoter, is flanked by loxP sites. Cre-mediated recombination during spermatogenesis of chimeric males removes the cassette leaving a single loxP site between exons 2-3. The construct was electroporated into C57BL/6-derived Bruce-4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts, and the resulting chimeric males were crossed to C57BL/6 females. Heterozygous b₇ (D146A) mice (harboring the D146A substitution, EcoRI insertion, and single remaining loxP site) were bred together for many generations prior to arrival at The Jackson Laboratory.

Control Information

	Control
	000664 C57BL/6J
Considerations for Choosing Controls

Related Strains

Strains carrying other alleles of Itgb7

002965	C57BL/6-Itgb7tm1Cgn/J
004944	NOD.B6-Itgb7tm1Cgn/2LtJ
004943	NOD.Cg-Selltm1Tft Itgb7tm1Cgn/LtJ

View Strains carrying other alleles of Itgb7     (3 strains)

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms

      assigned by genotype

Itgb7^tm1Mshi/Itgb7^tm1Mshi

        involves: C57BL/6

• immune system phenotype
• abnormal leukocyte migration/homing (MGI Ref ID J:127413)
  • labeled splenocytes have slightly less migration rate to the small and large intestine after intravenous injection into congenic hosts
  • migration is increased when splenocytes are pretreated with pertussis toxin
• abnormal leukocyte adhesion (MGI Ref ID J:127413)
  • splenocytes and memory T cells persistently exhibit firm adhesion to MadCAM coated surfaces even in presence of calcium ions that normally inhibit binding
  • five fold more splenocytes are found to be "sticking" to Peyer's patch venules
• colitis (MGI Ref ID J:127413)
  • only a mild form of colitis develops when naïve CD4 T cells from these mice are transferred to immunocompromised mice as opposed to the robust colitis that occurs when the T cells come from wild-type mice
  • the number of transferred T cells found in the large intestine is reduced by half
• small Peyer's patches (MGI Ref ID J:127413)
  • Peyer's patches are about 20% smaller in size
  • they contain four fold less T cells and three fold less B cells
• digestive/alimentary phenotype
• colitis (MGI Ref ID J:127413)
  • only a mild form of colitis develops when naïve CD4 T cells from these mice are transferred to immunocompromised mice as opposed to the robust colitis that occurs when the T cells come from wild-type mice
  • the number of transferred T cells found in the large intestine is reduced by half

View Research Applications

Research Applications

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

Cancer Research
Defects in Cell Adhesion Molecules

Cell Biology Research
Defects in Cell Adhesion Molecules

Developmental Biology Research
Defects in Cell Adhesion Molecules
Internal/Organ Defects (Lymphoid Tissue Defects)
Lymphoid Tissue Defects

Hematological Research
Immunological Defects (B and T cell deficiency)

Immunology and Inflammation Research
Immunodeficiency (B and T cell deficiency)
Immunodeficiency (B cell defects)
Immunodeficiency (B cell deficiency)
Immunodeficiency (Inflammatory bowel disease)
Immunodeficiency (T cell deficiency)
Inflammation (B and T cell deficiency)
Inflammation (Inflammatory bowel disease)
Lymphoid Tissue Defects (B and T cell deficiency)
Lymphoid Tissue Defects (Lymphocyte Homing)

Internal/Organ Research
Gastrointestinal Defects (colitis)
Lymphoid Tissue Defects (B and T cell deficiency)
Lymphoid Tissue Defects (T cell deficiency)

Research Tools
Cancer Research (B cell deficiency)
Cancer Research (T cell deficiency)
Immunology and Inflammation Research (B and T cell deficiency)
Immunology and Inflammation Research (B cell deficiency)
Immunology and Inflammation Research (T cell deficiency)

Virology Research
B and T Cell Deficiency

Genes & Alleles

Gene & Allele Information

Allele Symbol		Itgb7tm1Mshi
Allele Name		targeted mutation 1, Motomu Shimaoka
Allele Type		Targeted (knock-in)
Common Name(s)		Itgb7D146A;
Mutation Made By		Motomu Shimaoka,   Immune Disease Institute (formerly CBRI)
Strain of Origin		C57BL/6
ES Cell Line Name		Bruce 4
ES Cell Line Strain		C57BL/6
Gene Symbol and Name		Itgb7, integrin beta 7
Chromosome		15
Molecular Note		A vector consisting of a cre recombinase and a neomycin resistant gene under the control of the sperm-specific ACE promoter was used for targeting exon 4. In exon 4, the sequence encoding amino acid 146 was changed from an aspartate to an alanine (D146A). This mutation was predicted to keep the integrin in a constitutively adhesive state. The cre-recombinase and the neomycin resistance gene were excised in chimeras during spermatogenesis, leaving behind one loxP site. The mutation was confirmed by genomic sequencing. [MGI Ref ID J:127413]

Genotyping

Genotyping Information

Genotyping Protocols

Itgb7^tm1Mshi, STD PCR, vers. 1

Helpful Links

Optimizing PCR Protocols

References

References

Selected Reference(s)

Park EJ; Mora JR; Carman CV; Chen J; Sasaki Y; Cheng G; von Andrian UH; Shimaoka M. 2007. Aberrant activation of integrin alpha4beta7 suppresses lymphocyte migration to the gut. J Clin Invest 117(9):2526-38. [PubMed: 17786243]  [MGI Ref ID J:127413]

Health & husbandry

Health & Colony Maintenance Information

Animal Health Reports

Room Number           AX11

Colony Maintenance

Breeding & Husbandry	When maintaining a live colony, these mice may 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.