C.129S1(B6)-Nt5e^tm1Lft/J

Stock number: 027925
Product name: Cd73 KO on BALB/c
Research areas: Cancer Research, Cardiovascular Research, Immunology, Inflammation and Autoimmunity Research

Description

These mice may be useful when studying CD73-generated adenosine receptor signaling and antitumor immune responses.

Detailed description

A neo cassette replaces exon 3 of the ecto-5' nucleotidase (Nt5e) gene, abolishing CD73 enzyme activity. A shorter transcript is produced at about 10% of WT levels, but no functional protein is detectable. Nt5e encodes Cd73, a membrane-bound glycoprotein expressed on many cell types including subsets of T and B lymphocytes including regulator T cell (T_regs), endothelial cells, and mesenchymal cells. CD73 can hydrolyze nucleoside monophosphates, such as AMP, into bioactive intermediates like adenosine. CD73-generated adenosine activates transmembrane adenosine receptors and can act as either a pro- or anti-inflammatory mediator. Adenosine also regulates endothelial permeability, platelet activation, leukocyte adhesion, and lymphocyte migration into draining lymph nodes after an inflammatory stimulus. CD73-dependent adenosine receptor signaling protects mice during renal ischemia and inhibits vascular leakage during hypoxia. CD73 inhibits antitumor immune responses and is tolerogenic for cardiac and airway allografts. Homozygous CD73^-/- mice are viable and fertile. Under hypoxic conditions, CD73-deficient mice on the B6.129S1 background exhibit increased vascular leakage and display increased edema in all organs except the brain. CD73 deficient mice on the B6.129S1 background are protected from experimental autoimmune encephalomyelitis (EAE). The severity of graft-vs-host disease (GVHD) is increased in these mice, as are anti-tumor immune responses. These CD73-deficient mice on the congenic BALB/c background may be used with the 4T1 metastatic mammary tumor cell line.

In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. This is the case for the strain above. It should be noted that the phenotype could vary from that originally described. We will modify the strain description if necessary as published results become available.

Development

A targeting vector was designed to replace exon 3 of the ecto-5' nucleotidase, (Nt5e) gene with a neomycin resistance (neo) cassette in reverse orientation to the gene. The construct was electroporated into 129S1/Sv-Oca2⁺ Tyr⁺ Kitl⁺-derived CJ7 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric female was bred to a C57BL/6J male. The resulting heterozygous mice were crossed and their homozygous offspring were backcrossed to BALB/c mice for at least 14 generations. Upon arrival at The Jackson Laboratory, the mice were crossed to BALB/cByJ (001026) (Stock No. 001026) at least once to establish the colony.

Allele

Symbol: Nt5e^tm1Lft
Name: targeted mutation 1, Linda F Thompson
MGI accession ID: MGI:3522017
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Nt5e
Marker name: 5' nucleotidase, ecto
Chromosome: 9
Strain of origin: 129S1/Sv-Oca2⁺ Tyr⁺ Kitl⁺
Molecular note: Exon 3 was replaced with a neo cassette in reverse orientation to transcription. Northern blot of kidneys from mutant animlas indicated a lack of transcript.