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1 When indicated as verified ("YES"), then please note that information presented is to the best of our knowledge correct and up-to-date at the time of verification at the MMRRC Distribution Center; however, this information is subject to change due to breeding, maintenance, and other actions on the mouse strain at the MMRRC Distribution Center; direct any questions on this table to the MMRRC Distribution Center for this mouse stain.
2 If verification has not been performed (as indicated by "NO"), investigators may request specific verification testing for a fee. Requests should be submitted directly to the MMRRC Distribution Center assigned to the management, archiving, and distribution of the strain. A full listing of available testing and analytical services is available at https://www.mmrrc.org/about/services.php.
3 This information may or may not apply to each individual engineered allele (e.g., Cre, FlpO) present in the strain.
4 Recovery refers to thawing, in vitro fertilization (IVF), and/or embryo culture leading to live offspring.
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.
Conditional phenotype: No
To request gene-specific and other genotyping services for a strain, please contact the distribution MMRRC Center for more information.
The MMRRC has developed a Genetic Quality Control pipeline using the MiniMUGA array to provide additional information to identify and validate genetic backgrounds of MMRRC strains. For more information on whether genetic background data is available, please contact MMRRC_GeneticQC@med.unc.edu. Note: that MiniMUGA genetic background data is not available on all strains, but can be ordered if desired.
Disclaimer: If MMRRC Strain Genetic Quality Control (GQC; based on MiniMUGA genotyping and analysis) has been completed for this strain, the information might differ from the genetic background information provided by the submitter. MiniMUGA genetic analysis is done on a strain’s tissue samples taken when archived by or ordered from the assigned MMRRC Center.
Colony Surveillance Program and Current Health Reports
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.
A Commercial License Agreement from the Submitter is required for for-profit entities to use this strain. For more information, please contact Steven Tudor.
A Commercial License Agreement from the Submitter is required for for-profit entities to use this strain. For more information, please contact Steven Tudor
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.
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.
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