Strain Name:
C57BL/6J-Tg(APOA1-APOA1*)1Smijd/Mmmh
Stock Number:
065596-MU
Citation ID:
RRID:MMRRC_065596-MU
Other Names:
hAPOA1 4WF

Strain Information

APOA1
Name: apolipoprotein A1
Type: Gene
Species: Homo sapiens (human)
Chromosome:
Tg(APOA1-APOA1*)1Smijd
Name:
Type: Transgene
Species: Homo sapiens (human)
Chromosome:
Genetic Alterations
Site-directed mutagenesis was used to convert all four tryptophan residues in human apoA1to phenylalanine residues (TGG>TTT); see PMID:18688016), generating the 4WF-APOA1 isoform. Conversions were confirmed by Sanger sequencing.

HGVS nomenclature:
  • Genbank RefSeq - mRNA: NM_000039.2
  • Genbank RefSeq, protein: NP_000030.1
  • Variant, nucleic acid level: c.95_96delinsTT, c.221_222delinsTT, c.287_288delinsTT, c.395_396delinsTT
  • Variant, amino acid level, predicted: p.Trp32Phe, p.Trp74Phe, p.Trp96Phe, p.Trp132Phe
  • Check these variants, LUMC Mutalyzer: Sub#1, Sub#2, Sub#3, Sub#4
Note: The apoA1 protein is produced as a pre-protein, which is cleaved twice to release the mature human apoA1 isoform.
It is the consensus in the apoA1 field to number the residues vs. the mature 243-aa protein. Publication PMID:18688016 places to the Trp residues mentioned above at positions 8, 50, 72, and 108 in reference to sequence GenBank:CAA00975.1.
Genotype Determination
Phenotype
Male mice had significantly higher plasma levels than females for both isoforms of human APOA1, attributed to different production rates. With matched plasma levels, 4WF transgenics had a trend for slightly less HDL cholesterol vs. human APOA1 transgenics. While 4WF transgenics had 31% less reverse cholesterol transport (RCT) to the plasma compartment, equivalent RCT to the liver and feces were observed. Plasma from both strains had similar ability to accept cholesterol and facilitate ex-vivo cholesterol efflux from macrophages. 4WF-transgenic HDL was partially (~50%) protected from MPO-mediated loss of function while human APOA1-transgenic HDL lost all ABCA1-dependent cholesterol acceptor activity.
MeSH Terms
  • ATP Binding Cassette Transporter 1/genetics
  • ATP Binding Cassette Transporter 1/metabolism
  • Animals
  • Apolipoprotein A-I/genetics
  • Apolipoprotein A-I/metabolism
  • Cholesterol, HDL/genetics
  • Cholesterol, HDL/metabolism
  • Female
  • Humans
  • Macrophages/metabolism
  • Male
  • Mice
  • Mice, Transgenic
  • Oxidation-Reduction
  • Peroxidase/genetics
  • Peroxidase/metabolism
  • Protein Isoforms/genetics
  • Protein Isoforms/metabolism
  • Structure-Activity Relationship
  • ATP Binding Cassette Transporter 1
  • ATP-Binding Cassette Transporters/metabolism
  • Amino Acid Substitution
  • Apolipoprotein A-I/blood
  • Apolipoprotein A-I/chemistry
  • Apolipoprotein A-I/isolation & purification
  • Atherosclerosis/enzymology
  • Atherosclerosis/metabolism
  • Cell Line
  • Cholesterol/metabolism
  • Hydrogen Peroxide/metabolism
  • Lysine
  • Methionine
  • Mutagenesis, Site-Directed
  • Recombinant Proteins/metabolism
  • Tryptophan
Strain Development
Subcloned a 12.3-kb EcoRI fragment into the EcoRI site of pUC19, derived from a human BAC clone (CTD-2149o15, Invitrogen) that contained the entire APOA1 gene, including 6.3 and 3.7 kb of 5' and 3'-flanking regions, respectively.

The modified EcoRI insert was microinjected into C57BL/6-fertilized oocytes (Case Transgenic Core lab). Transgenic founders 4WF-APOA1 were obtained, screened for the human plasma APOA1 levels by dot blot, and the highest expressing line was selected for further breeding (sib-mating) and study.
Suggested Control Mice
C57BL/6J and C57BL/6-Tg(APOA1)1Rub/J mice (JAX Strain #001927)
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@missouri.edu. Older strains may not have this information.
  • Cardiovascular
Donor
Jonathan Smith, Ph.D., Cleveland Clinic.
Primary Reference

Berisha SZ, Brubaker G, Kasumov T, Hung KT, DiBello PM, Huang Y, Li L, WillardB, Pollard KA, Nagy LE, Hazen SL, Smith JD. HDL from apoA1 transgenic mice expressing the 4WF isoform is resistant to oxidative loss of function. J LipidRes. 2015 Mar;56(3):653-64. doi: 10.1194/jlr.M056754. Epub 2015 Jan 5. PubMedPMID: 25561462; PubMed Central (Medline PMCID: PMC4340312)

Peng DQ, Brubaker G, Wu Z, Zheng L, Willard B, Kinter M, Hazen SL, Smith JD. Apolipoprotein A-I tryptophan substitution leads to resistance to myeloperoxidase-mediated loss of function. Arterioscler Thromb Vasc Biol. 2008 Nov;28(11):2063-70. doi: 10.1161/ATVBAHA.108.173815. Epub 2008 Aug 7. (Medline PMID: 18688016)

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@missouri.edu.
Coat Color
Black
Eye
Black
MMRRC Breeding System
Sib-mating
Generation
F10+
Overall Breeding Performance
Good
NOTE: "Hemizygote" as used here refers to males carrying a mutation on the X Chromosome or mice of either sex carrying an inserted transgene with no homologous allele on the other chromosome.
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Yes Yes
Hetero/Hemizygotes are fertile: Yes Yes
Age Reproductive Decline: 10 to 12 months 10 to 12 months
Average litter size
4 to 6
Recommended wean age
3 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 mmrrc@missouri.edu 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
065596-MU-SPERM Cryo-preserved spermatozoa $437.00 / Non-Profit Aliquot Approximate quantity3
065596-MU-RESUS Litter recovered from cryo-archive $2,624.00 / Non-Profit Litter Recovered litter4; additional fees for any special requests.
Cryopreserved material may be available upon request, please inquire to mmrrc@missouri.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.