Description

Strain Information

Former Names B6.129S6-Eraftm1Mjwe/J    (Changed: 05-FEB-08 ) Type Congenic; Mutant Strain; Targeted Mutation; Additional information on Genetically Engineered Mutant Mice. Mating System Heterozygote x Heterozygote         (Female x Male) Species laboratory mouse Generation N10+ (12-SEP-08)   Donating Investigator Mitchell Weiss,   Children's Hospital of Philadelphia

Description
Mice homozygous for this targeted allele (AHSP^-/- or ERAF^-/-) are fertile with normal lifespans up to at least 18 months of age. No RNA or protein from the targeted gene is detected in hematopoietic tissues from homozygotes. AHSP^-/- mice exhibit abnormal erythrocyte morphology with intracellular inclusion bodies that stain positively for denatured hemoglobin (Heinz bodies). Homozygous mice also have reduced lifespan of circulating red blood cells, increased apoptosis of erythroid precursors, and increased production of reactive oxygen species (ROS) with consequent damage to hemoglobin A and other cellular components. As the a-hemoglobin stabilizing protein specifically binds the cytotoxic free a-Hb subunit of hemoglobin A, these AHSP-mutant mice may be useful in studying erythroid development/erythropoiesis, thalassemia, Heinz body hemolytic anemia, and other hemoglobinopathies.

In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. As AHSP-mutant mice were originally described on a mixed C57BL/6 and 129 genetic background, it should be noted that the phenotype could vary from that originally described. We will modify the strain description if necessary as published results become available.

Development
A targeting vector designed to replace the entire coding sequence of the targeted gene with a loxP-flanked pgk-neo cassette was electroporated into 129S6/SvEvTac-derived TL1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimera's were bred with C57BL/6 mice to produce heterozygous mutants. Mice were then bred to mice with cre expression in sperm (undetermined genetic background) to remove the neo cassette. The resulting offspring (with a single loxP site replacing the entire coding region) were selected. This mutant strain was then backcrossed to C57BL/6 bred for at least 9 generations prior to arrival at The Jackson Laboratory.

Control Information

	Control
	Wild-type from the colony
	000664 C57BL/6J
Considerations for Choosing Controls

Additional Web Information

Congenic Nomenclature

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms

      assigned by genotype

The following phenotype information may relate to a genetic background differing from this JAX^® Mice strain.

Eraf^tm1.1Mjwe/Eraf⁺

        involves: 129S6/SvEvTac * C57BL/6

• hematopoietic system phenotype
• abnormal erythrocyte morphology (MGI Ref ID J:94421)
  • erythrocytes also contain inclusion bodies composed of denatured hemoglobin chains (Heinz bodies)

Eraf^tm1.1Mjwe/Eraf^tm1.1Mjwe

        involves: 129S6/SvEvTac * C57BL/6

• hematopoietic system phenotype
• abnormal hematopoiesis (MGI Ref ID J:94421)
  • there is an elevated level of apoptotic erythroid precursors in mutant spleens
• abnormal erythrocyte morphology (MGI Ref ID J:94421)
  • target cells (codocytes) and spiculated cells can be seen in blood smears
  • some erythrocytes have inclusion bodies composed of denatured hemoglobin chains (Heinz bodies)
  • erythrocytes exhibit a half-life of 12 days, compared to 22 days of cells from wild-type littermates
  • erythroid precursors show significant hyperplasia in the spleen and to a lesser degree in the bone marrow
• anisopoikilocytosis (MGI Ref ID J:94421)
  • blood smears show morphologic abnormalities such as irregular size and shape
• anisocytosis (MGI Ref ID J:94421)
  • significant variation in size of erythrocytes as shown by increased red blood cell distribution width (RBW)
• decreased mean corpuscular volume (MGI Ref ID J:94421)
  • mice have small red blood cells with low mean corpuscular hemoglobin (13.6 pg vs 16.2 pg in wild-type)
• polychromatophilia (MGI Ref ID J:94421)
  • large inclusions are present only in polychromatophilic cells, suggesting accelerated erythrocyte destruction
• abnormal hemoglobin (MGI Ref ID J:94421)
  • erythrocytes contain alpha- and beta-globin precipitates; membrane-associated globin chains are not observed in control littermates
• abnormal hemoglobin content (MGI Ref ID J:94421)
• increased hemoglobin concentration distribution width (MGI Ref ID J:94421)
  • variation in hemoglobin content of mutant erythrocytes, evidenced by increased hemoglobin distribution width (HDW), is observed
• decreased mean corpuscular hemoglobin (MGI Ref ID J:94421)
• anemia (MGI Ref ID J:94421)
  • mice show mild anemia associated with small red blood cells
• decreased hematocrit (MGI Ref ID J:94421)
  • hematocrit is reduced to 48.7% from 55.8% in wild-type
• increased erythroid progenitor cell number (MGI Ref ID J:94421)
  • spleen shows increased proportion of erythroid precursors
  • cultured spleen cells show a higher proportion of burst-forming unit erythroid and colony-forming unit erythroid cells (BFUe and CFUe's)
• reticulocytosis (MGI Ref ID J:94421)
  • reticulocyte count is elevated (to 4.4% from 2.2% in wild-type)
• enlarged spleen (MGI Ref ID J:94421)
  • spleen is significantly enlarged
• cellular phenotype
• increased cellular sensitivity to oxidative stress (MGI Ref ID J:94421)
  • mutants show a larger decrease in hematocrit after phenylhydrazine treatment resulting from hemolytic anemia compared to control littermates
• oxidative stress (MGI Ref ID J:94421)
  • mutant erythrocytes display intrinsically elevated oxidative stress
  • mice have increased levels of reactive oxygen species catalyzed by alpha-hemoglobin
• immune system phenotype
• enlarged spleen (MGI Ref ID J:94421)
  • spleen is significantly enlarged

View Research Applications

Research Applications

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

Developmental Biology Research
Lymphoid Tissue Defects (hematopoietic defects)

Hematological Research
Anemia, Iron Deficiency and Transport Defects
Anemia, Iron Homeostasis Defects
Hematopoietic Defects
Hemoglobin Defects (beta^O-thalassemia)
Hemoglobin Defects (thalassemia)

Research Tools
Hematological Research

Genes & Alleles

Gene & Allele Information

Allele Symbol		Eraftm1.1Mjwe
Allele Name		targeted mutation 1.1, Mitchell J Weiss
Allele Type		Targeted (knock-out)
Common Name(s)		AHSP-/-;
Mutation Made By		Mitchell Weiss,   Children's Hospital of Philadelphia
Strain of Origin		129S6/SvEvTac
ES Cell Line Name		TL1/TL-1
ES Cell Line Strain		129S6/SvEvTac
Gene Symbol and Name		Eraf, erythroid associated factor
Chromosome		UN
Gene Common Name(s)		AHSP; EDRF;
Molecular Note		A targeting vector was designed to replace the entire coding sequence of the targeted gene with a loxP-flanked pgk-neo cassette. Mice were then bred to spermadine-cre mice to remove the neo cassette. The resulting offspring have a single loxP site replacing the entire coding region. [MGI Ref ID J:77038] [MGI Ref ID J:94421]

Genotyping

Genotyping Information

Genotyping Protocols

Eraf^tm1.1Mjwe, STD PCR, vers. 1

Helpful Links

Optimizing PCR Protocols

References

References

Selected Reference(s)

Kihm AJ; Kong Y; Hong W; Russell JE; Rouda S; Adachi K; Simon MC; Blobel GA; Weiss MJ. 2002. An abundant erythroid protein that stabilizes free alpha-haemoglobin. Nature 417(6890):758-63. [PubMed: 12066189]  [MGI Ref ID J:77038]

Kong Y; Zhou S; Kihm AJ; Katein AM; Yu X; Gell DA; Mackay JP; Adachi K; Foster-Brown L; Louden CS; Gow AJ; Weiss MJ. 2004. Loss of alpha-hemoglobin-stabilizing protein impairs erythropoiesis and exacerbates beta-thalassemia. J Clin Invest 114(10):1457-66. [PubMed: 15545996]  [MGI Ref ID J:94421]

Yu X; Kong Y; Dore LC; Abdulmalik O; Katein AM; Zhou S; Choi JK; Gell D; Mackay JP; Gow AJ; Weiss MJ. 2007. An erythroid chaperone that facilitates folding of alpha-globin subunits for hemoglobin synthesis. J Clin Invest 117(7):1856-65. [PubMed: 17607360]  [MGI Ref ID J:124209]

Health & husbandry

Health & Colony Maintenance Information

Animal Health Reports

Room Number           AX11

Colony Maintenance

Breeding & Husbandry	When maintaining a live colony, homozygous mice may be bred.
Mating System	Heterozygote x Heterozygote         (Female x Male)

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
	Wild-type from the colony
	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.

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Ordering and Purchasing Information

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Contact Information
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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.”

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