NOD.Cg-Prkdc^scid Il2rg^tm1Wjl Ace2^em1Lutzy/J

Stock number: 035424
Product name: NSG Ace2^H353K
Research areas: Cancer Research, Developmental Biology Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

NSG Ace2^H353K knockin mice express an H353K (histidine to lysine) amino acid substitution in the Ace2 (angiotensin I converting enzyme (peptidyl-dipeptidase A) 2) locus on the immunodeficient NSG background (NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ, Stock No. 005557). 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).

This strain may be useful for the study of 2019 novel coronavirus (SARS-CoV-2) pathogenesis as well as cardiovascular disease.

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.

Stock No. 035424 NSG Ace2^H353K is a multiple mutant strain created by CRISPR/cas9 genome editing of the Ace2 locus in NSG mice (Stock No. 005557). In an attempt to offer popular alleles together, mouse lines may be bred together for purpose of researchers to study and characterize. It should be noted that the phenotype of Stock No. 035424 could vary from that originally described for the individual mouse lines used to assemble it. We may modify the strain description if necessary as published results become available. Each mutant allele is briefly described below.

The Ace2^H353K knock-in allele was created using CRISPR/cas9 genome editing to express the H353K (histidine to lysine) amino acid substitution in the mouse 353 codon of the Ace2 (angiotensin I converting enzyme (peptidyl-dipeptidase A) 2) locus on the X chromosome. Ace2 encodes a dipeptidyl carboxydipeptidase involved in cleavage of angiotensin I and II. Mouse Ace2 has high homology to human ACE2. 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. A lysine is present at position 353 of ACE2 in humans, non-human primates, bats and ferrets, while mice and rats have a histidine at this position. In mice, the Ace2 H353K mutation has been found to confer susceptibility to SARS-CoV-2 infection in vivo - see Stock No. 034903.

The NSG component involves two mutations on the NOD/ShiLtJ genetic background - severe combined immune deficiency (scid) and a complete null allele of the IL2 receptor common gamma chain (Il2rg^null). The scid mutation is in the DNA repair complex protein Prkdc and renders the mice B and T cell deficient. The IL2rg^null mutation prevents cytokine signaling through multiple receptors, leading to a deficiency in functional NK cells. The severe immunodeficiency allows the mice to be humanized by engraftment of human CD34+ hematopoietic stem cells (HSC), peripheral blood mononuclear cells (PBMC), patient derived xenografts (PDX), or adult stem cells and tissues. The immunodeficient NSG mice enable research in human immune function, infectious disease, diabetes, oncology, and stem cell biology.

Females homozygous for all three alleles and males homozygous for Prkdc and hemizygous for the Il2rg^tm1Wjl and Ace2^em1Lutzy alleles are viable and fertile - please note that Il2rg and Ace2 are X-linked genes in cis (on the same chromosome) ~63 Mbp apart.

Development

The Ace2^H353Kallele was created by Dr. Cathy Lutz (The Jackson Laboratory) using constructs designed by Dr. Paul B. McCray (University of Iowa). CRISPR/cas9 genome editing was used to create an amino acid substitution at position 353 (histidine to lysine, H353K) in the Ace2 locus on the X chromosome. Single guide RNA and cas9 endonuclease were introduced into NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ -derived fertilized eggs by pronuclear injection. Embryos were transferred to pseudopregnant females, and correctly targeted pups were bred to NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ (NSG), Stock No. 005557, for germline transmission. The Il2rg and Ace2 mutations are in cis (on the same chromosome) ~63 Mbp apart. The colony was maintained on the NSG background.

Allele

Symbol: Ace2^em1Lutzy
Name: endonuclease-mediated mutation 2, Cathy Lutz
MGI accession ID: MGI:6865644
Generation method: Endonuclease-mediated
Attributes: Humanized sequence
Synonyms: Ace2^H353K
Marker symbol: Ace2
Marker name: angiotensin I converting enzyme (peptidyl-dipeptidase A) 2
Marker synonyms: 2010305L05Rik; ACEH
Chromosome: X
Strain of origin: NOD.Cg-Prkdc^scid Il2rg^tm1Wjl/SzJ
Molecular note: CRISPR/cas9 genome editing is used to create an amino acid substitution at position 353 (histidine to lysine, H353K). A lysine is present at position 353 of ACE2 in humans, non-human primates, bats and ferrets, while mice and rats have a histidine at this position.

Allele

Symbol: Prkdc^scid
Name: severe combined immunodeficiency
MGI accession ID: MGI:1857113
Generation method: Spontaneous
Synonyms: scid
Marker symbol: Prkdc
Marker name: protein kinase, DNA activated, catalytic polypeptide
Marker synonyms: DNAPDcs; DNAPK; DNA-PK; DNAPKc; DNA-PKC; DNA-PKcs; DNPK1; DOXNPH; dxnph; HYRC; HYRC1; IMD26; p350; S473K; slip; XRCC7
Chromosome: 16
Strain of origin: C.BKa-Igh^b/Icr
Site of expression: T and B lymphocytes.
Molecular note: A T-to-A transversion point mutation at a position corresponding to codon 4046 (codon 4095 in transcript ENSMUST00000023352.8) created a premature stop codon (p.Y4046*).

Allele

Symbol: Il2rg^tm1Wjl
Name: targeted mutation 1, Warren J Leonard
MGI accession ID: MGI:1857455
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: [KO]gammac; CD132^-; gammac^-; gc^-; Il2rg^tm1Wjll; IL2Rgamma^null
Marker symbol: Il2rg
Marker name: interleukin 2 receptor, gamma chain
Marker synonyms: [g]c; Ab2-183; Cd132; CIDX; common cytokine receptor gamma chain; common gamma chain; gamma C receptor; gamma(c); gammaC; gc; IL-2RG; IMD4; P64; SCIDX; SCIDX1
Chromosome: X
Strain of origin: 129S4/SvJae
Site of expression: Primarily lymphoid cells.
Molecular note: A neomycin resistance cassette replaced part of exon 3 and all of exons 4 - 8 of the gene, resulting in the loss of most of the extracellular domain and all of the transmembrane and cytoplasmic domains of the protein.