NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl Tg(SLC10A1)15Mvw/MvwJ

Stock number: 030533
Product name: hNTCP/BAC-NRG
Research areas: Cancer Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

hNTCP/BAC-NRG mice express human SLC10A1 (or NTCP) on an immunodeficient background and are susceptible to infection with hepatitis delta virus (HDV). They may be used as a humanized mouse model for hepatitis delta virus infection.

Detailed description

SLC10A1 (solute carrier family 10 (sodium/bile acid cotransporter family), member 1 or NTCP) is an integral membrane glycoprotein that is found in the basolateral membranes of hepatocytes. SLC10A1 is involved in the reabsorption of bile acids and acts as the functional receptor for hepatitis delta virus (HDV) and hepatitis B virus (HBV). Mice carrying the hNTCP/BAC transgene express part of human chromosome 14 including SLC10A1 (or NTCP) and the upstream human regulatory elements. Transgene expression is observed in the liver, but not in kidney, brain, lung or intestine. Infection with HDV is dependent on the presence of HBsAg. On the immunodeficient NRG background (NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl/SzJ, Stock No. 007799), hNTCP/BAC-NRG mice treated with 1/3x over length of HBV genome and then infected with HDV developed a persistent HDV infection for more than 80 days. Although HBV-challenged hNTCP/BAC-NRG mice exhibit HBV in 25% of hepatocytes 18 days post-challenge, there is no evidence of productive HBV infection.

Immunocompetent mice carrying only the human SLC10A1 (or NTCP) transgene (hNTCP/BAC) are available as Stock No. 030535.

Similar to other immunodeficient strains, maintaining homozygous NRG mice in high health status (specific pathogen-free) vivaria promotes overall colony health. If homozygous animals are maintained in low health barrier rooms, the use of medicated water (e.g., sulfatrim/trimethoprim-sulfa or enrofloxacin/Baytril) is suggested to increase overall colony health.

Development

BAC RP11-72J8 was modified to include a 982bp synthetic gene that includes 500 bp of ROSA26 sequence to allow in vivo linearization by the CRISPR/Cas9 complex. Specifically, the gene contains from 5’ to 3’: human SLC10A1 sequence, an FRT-flanked PGK-Neo cassette, 500 bp of ROSA26 sequence and human SLC10A1 sequence (SLC10A1 sequence totals 397 bp). Sequence analysis indicated that the ROSA26 sequence is 76,428 bp upstream of exon 1 of the SLC10A1 gene. Single guide RNAs were designed to target the ROSA26 locus and the RP11-72J8 BAC. The sgRNAs and Cas9 endonuclease were introduced into NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl/J-derived (NRG - Stock No. 007799) fertilized eggs by pronuclear injection. Embryos were transferred to pseudopregnant females, and correctly targeted pups were bred to NRG mice to verify germline transmission. Founder 15 was selected to establish this line. Sequence analysis indicated that the transgene did not insert in the ROSA26 locus.

Please note, breeding results suggest that the transgene may be X-linked.

Allele

Symbol: Rag1^tm1Mom
Name: targeted mutation 1, Peter Mombaerts
MGI accession ID: MGI:1857241
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: Rag-; Rag^1tm1Mom; RAG-1-; Rag1^-; RAG1^null; Rag-1KO
Marker symbol: Rag1
Marker name: recombination activating gene 1
Marker synonyms: RAG-1; RNF74
Chromosome: 2
Strain of origin: 129S7/SvEvBrd-Hprt1⁺
Site of expression: expression is seen in bone marrow derived cell lines.
Molecular note: A 1356 bp genomic fragment of the Rag1 gene, encoding the nuclear localization signal and the zinc-finger motif, was replaced by a neomycin cassette. A mutant transcript expressed from this allele was detected by Northern blot in bone marrow derived cell lines from homozygous mice.

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.

Allele

Symbol: Tg(SLC10A1)15Mvw
Name: transgene insertion 15, Micheal Wiles
MGI accession ID: MGI:5883447
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: hNTCP/BAC-NRG
Marker symbol: Tg(SLC10A1)15Mvw
Marker name: transgene insertion 15, Micheal Wiles
Chromosome: UN
Strain of origin: NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl/SzJ
Site of expression: The transgene is expressed in the liver, but not in kidney, brain, lung or intestine.
Molecular note: BAC RP11-72J8 was modified to include a 982bp synthetic gene that includes 500 bp of ROSA26 sequence to allow in vivo linearization by the CRISPR/Cas9 complex. Specifically, the gene contains from 5' to 3': human SLC10A1 sequence, an FRT-flanked PGK-Neo cassette, 500 bp of ROSA26 sequence and human SLC10A1 sequence (SLC10A1 sequence totals 397 bp). Sequence analysis indicates that the ROSA26 sequence is 76,428 bp upstream of exon 1 of the SLC10A1 gene. Single guide RNAs are designed to target the ROSA26 locus and the RP11-72J8 BAC.

Allele

Symbol: rd24
Name: retinal degeneration 24
MGI accession ID: MGI:7643884
Generation method: Spontaneous
Attributes: Not Specified
Marker symbol: rd24
Marker name: retinal degeneration 24
Chromosome: 5
Strain of origin: NOD.Cg-Rag1^tm1Mom Il2rg^tm1Wjl/SzJ
Molecular note: This spontaneous heritable phenotypic mutation has been mapped to Chromosome 5 closely linked to D5Mit356 with no crossovers in 100 F2 mice. Sequence characterization has not been done but the most likely candidate gene is Cnga1.