C57BL/6J-Ace2^em1Guru/Mmjax

Stock number: 036479
Product name: B6.mAce2KI-31K,82M,353K
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

mAce2KI-31K,82M,353K knock-in mice contain five amino acid substitutions (initially reported as, N31K, S82M, H353K, plus N30D and Q34K) in the Ace2 (angiotensin I converting enzyme (peptidyl-dipeptidase A) 2) gene. Ace2 is the receptor used for cellular entry by several coronaviruses, including severe acute respiratory syndrome coronavirus-1 (SARS-CoV) and -2 (SARS-CoV-2). These mutations function to humanize the mouse Ace2 coding sequence by altering it to match human coding sequence in a region important for binding to the SARS-CoV-2 spike protein. This strain may be useful for the study of 2019 novel coronavirus (SARS-CoV-2) pathogenesis.

Detailed description

Please note, these mice should be handled in a manner consistent with CDC/ABSA/WHO guidelines for prevention of human infection with the SARS-CoV-2 virus. Proper PPE and handling methods should be used at all times when working with these mice. Additional important guidelines for using SARS-CoV-2 susceptible mouse lines.

Ace2 (angiotensin I converting enzyme (peptidyl-dipeptidase A) 2) encodes a dipeptidyl carboxydipeptidase that functions to catalyze cleavage of angiotensin I and II. Human ACE2 is the receptor used for cellular entry by several coronaviruses, including severe acute respiratory syndrome coronavirus-1 (SARS-CoV), severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2; 2019 novel coronavirus) and HCoV-NL63. mAce2KI-31K,82M,353K knock-in mice contain five amino acid substitutions (initially reported as, N31K, S82M, H353K, plus N30D and Q34K) designed to completely humanize the mouse Ace2 coding sequence in a region important for binding to the SARS-CoV-2 spike protein. Mice homozygous (or hemizygous) for the mutation are viable and fertile. At this time, June 2021, susceptibility to virus has not been tested. This strain may be useful for the study of 2019 novel coronavirus (SARS-CoV-2) pathogenesis.

Development

The mAce2KI-30D,31K,34D,82M,353K knock-in allele was created by Dr. Channabasavaiah Gurumurthy at the University of Nebraska Medical Center. CRISPR/cas9 endonuclease-mediated genome editing was used to insert 5 amino acid substitutions located in three different exons of the Ace2 (angiotensin I converting enzyme (peptidyl-dipeptidase A) 2) gene on the X Chromosome. The amino acids 30, 31, 34 in exon 2 were changed from asparagine to glutamic acid (N30D), asparagine to lysine (N31K), and glutamine to aspartic acid (Q34D) respectively, amino acid 82 in exon 3 was changed from serine to methionine (S82M), and the amino acid 353 in exon 9 was changed from histidine to lysine (H353K). Four single guide RNAs and Cas9 endonuclease were injected into C57BL/6J-derived fertilized eggs by pronuclear injection. Embryos were transferred to pseudopregnant females, and correctly targeted pups were bred to C57BL/6J mice for germline transmission. Sequence was confirmed by Sanger sequencing; off-target screening has not been performed. The colony was maintained on the C57BL/6J background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

Allele

Symbol: Ace2^em1Guru
Name: endonuclease-mediated mutation 1, Channabasavaiah Gurumurthy
MGI accession ID: MGI:6763023
Generation method: Endonuclease-mediated
Attributes: Humanized sequence
Synonyms: mAce2^{KI-31K,82M,353K}
Marker symbol: Ace2
Marker name: angiotensin I converting enzyme (peptidyl-dipeptidase A) 2
Marker synonyms: 2010305L05Rik; ACEH
Chromosome: X
Strain of origin: C57BL/6J
Molecular note: CRISPR/cas9 endonuclease-mediated genome editing is used to insert 5 amino acid substitutions. The amino acids 30, 31, 34 in exon 2 were changed from asparagine to glutamic acid (N30D), asparagine to lysine (N31K), and glutamine to aspartic acid (Q34D) respectively, amino acid 82 in exon 3 was changed from serine to methionine (S82M), and the amino acid 353 in exon 9 was changed from histidine to lysine (H353K). The mutations are designed to completely humanize the mouse Ace2 coding sequence in a region important for binding to the SARS-CoV-2 spike protein.