Loading Mouse GIF
Loading...

Strain Name:
C57BL/6N-Ace2em1(ACE2)MbpTmprss2em1(TMPRSS2)MbpFurinem1(FURIN)Mbp/Mmucd
Stock Number:
071887-UCD
Citation ID:
RRID:MMRRC_071887-UCD
Other Names:
C57BL/6N-Ace2em1(ACE2)MbpTmprss2em1(TMPRSS2)MbpFurinem1(FURIN)Mbp/Mmucd

Strain Information

Ace2em1(ACE2)Mbp
Name: angiotensin converting enzyme 2; endonuclease-mediated mutation 1, Mouse Biology Program, UC Davis
Type: Allele
Species: Multi-species
Chromosome: X
Alteration at locus: CRISPR
Tmprss2em1(TMPRSS2)Mbp
Name: transmembrane protease, serine 2; endonuclease-mediated mutation 1, Mouse Biology Program, UC Davis
Type: Allele
Species: Multi-species
Chromosome: 16
Alteration at locus: CRISPR
Furinem1(FURIN)Mbp
Name: furin, paired basic amino acid cleaving enzyme; endonuclease-mediated mutation 1, Mouse Biology Program, UC Davis
Type: Allele
Species: Multi-species
Chromosome: 7
Alteration at locus: CRISPR
Furin
Name: furin, paired basic amino acid cleaving enzyme
Synonyms: Fur, PACE, SPC1, Pcsk3, 9130404I01Rik
Type: Gene
Species: Mouse
Chromosome: 7
Alteration at locus: CRISPR
NCBI: 18550
HGNC: HGNC:8568
Homologene: 1930
Ace2
Name: angiotensin converting enzyme 2
Synonyms: 2010305L05Rik
Type: Gene
Species: Mus musculus (mouse)
Chromosome: X
Alteration at locus: CRISPR
NCBI: 70008
Homologene: 41448
Tmprss2
Name: transmembrane protease, serine 2
Synonyms: epitheliasin, D16Ertd61e
Type: Gene
Species: Mouse
Chromosome: 16
Alteration at locus: CRISPR
NCBI: 50528
VEGA: 16
Homologene: 4136
Genetic Alterations

Human ACE2 DNA consisting of 5' UTR, CDS (exons 2-19) fused to mouse Ace2 3'UTR replaced the DNA region in mouse Ace2 gene stretching from the 5'UTR to the starting ATG in exon 2. The approach minimizes potential downstream splicing or RNP re-cleavage. Expression of the human CDS is primarily controlled by the mouse regulatory system and is expected to not transcribe the mouse version. Gene expression showed expected human ACE2 levels whereas mouse Ace2 was not expressed. CRISPR RNP was used to assist with homologous recombination using dsDNA repair in JM8A3 ES cells.


Human TMPRSS2 exons 3-14 (ENSG00000184012.14) was inserted in-frame into exon 2 (ENSMUSE00000641779) of the mouse Tmprss2 gene. CRISPR RNP was used to assist with homologous recombination (HR) using dsDNA repair template in ES cells. The mouse exon 2 splice acceptor and the guide protospacer (nt 1-17) were removed from the HR template to prevent potential downstream splicing or RNP re-cleavage. The expression of the human CDS is controlled by the mouse regulatory systems (including 3’UTR), and predicted to terminate with the transgene’s STOP codon without expression of the endogenous sequence. Gene expression showed expected human TMPRSS2 levels whereas mouse Tmprss2 was not expressed. CRISPR RNP was used to assist with homologous recombination using dsDNA repair template in JM8A3 ES cells derived from C57BL/6N.


Human FURIN DNA consisting of CDS (exons 2-16) fused to mouse Furin 3'UTR replaced start codon in mouse Furin gene. This approach minimizes potential downstream splicing or RNP re-cleavage as the guide protospacer was split. Expression of the human CDS is primarily controlled by the mouse regulatory system and is expected to not transcribe the mouse version. CRISPR RNP was used to assist with homologous recombination using dsDNA repair in JM8A3 ES cells.


For each of the three lines above, following blastocyst injection, the chimeras were mated with C57BL/6NCrl to create germline heterozygotes. Long-range PCR products were fully sequenced; potential off-targets were absent. Subsequent backcross was to C57BL/6NCrl.

Genotype Determination
ES Cell Line
JM8A3
Phenotype
This model replaces the expression of the mouse Ace2, Tmprss2 and Furin genes with expression of the human ACE2, TMPRSS2 and FURIN genes. Other than expression of the human genes and inhibition of expression of the corresponding murine genes, this mouse exhibits no known observable or measurable phenotype that differs from the genetic background wildtype strain (C57BL/6N). However, it is anticipated that once exposed to SARS-CoV-2 virus (10^5), these mice will exhibit signs of disease that are not seen in wildtype mice.

Conditional phenotype: No

Strain Development
For each of the three alterations, CRISPR RNP was used to assist with homologous recombination (HR) using dsDNA repair template in JM8A3 ES cells. Following blastocyst injection, the chimeras were mated with C57BL/6NCrl to create germline heterozygotes. Long-range PCR products were fully sequenced; potential off-targets were absent. Subsequent backcross was to C57BL/6NCrl. Each of the ACE2, TMPRSS2, and FURIN mouse lines were then intercrossed to create a homozygous tri-genic ACE2/TMPRSS2/FURIN mouse model.
Suggested Control Mice
C57BL/6NCrl or wild-type siblings
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@ucdavis.edu. Older strains may not have this information.
  • Immunology and Inflammation
  • Models for Human Disease
Donor
Kent Lloyd, D.V.M., University of California, Davis.
Primary Reference
Not ready to publish

Colony and Husbandry Information

Colony Surveillance Program and Current Health Reports

For more information about this colony's health status contact mmrrc@ucdavis.edu
Coat Color
Black
Eye
Black
MMRRC Breeding System
Sib-mating
Generation
Unknown
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 X-linked
Homozygotes are fertile: Yes X-linked
Hetero/Hemizygotes 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 (up to 2 males and 2 females or 4 mice) per investigator per month are available from a live colony, usually available to ship in under 12 weeks. Larger quantities may be available, please contact the distributing center directly at mmrrc@ucdavis.edu for more details.

Cryopreserved material may be available upon request, please inquire to mmrrc@ucdavis.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.

- Products for this strain are Not Yet Available for Ordering
- If you register interest in this strain, you will be notified when it becomes available for ordering.

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.