Cloning, sequencing and expression analysis of the mouse NOD2/CARD15 gene

Y. Iwanaga, Michael Davey, Tammy Martin, Stephen Planck, M. L. DePriest, M. M. Baugh, C. M. Suing, James (Jim) Rosenbaum

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Background: Mutations in the human NOD2/CARD15 gene have been associated with Crohn's disease and Blau syndrome. The objective of the present study was to clone the murine form of NOD2 and characterize its tissue distribution, function and response to inflammatory stimuli. Methods: Murine NOD2 was isolated using anchored polymerize chain reaction (PCR). Sequence analysis confirmed the identification of full-length cDNA representing the murine NOD2 gene. Using this sequence to search a Mus musculus supercontig database, NOD2 genomic DNA was identified. NOD2 was transfected into human embryonic kidney (HEK) cells and nuclear factor kappa B (NF-κB) activation was measured using a reporter assay. Tissue distribution and changes in transcription in mouse monocytes in response to inflammatory stimuli was determined by real time PCR. Results: The NOD2 gene spans 39 KB and contains 12 coding exons on chromosome 8. Expression of mouse NOD2 into HEK cells resulted in NF-κB activation. NOD2 was found to be expressed in all mouse tissues analyzed except skin, with highest levels in lung, thymus and spleen. NOD2 mRNA levels increased greater than two-fold in a monocyte cell line in response to lipopolysaccharide, lipoteichoic acid, interferon-γ and tumor necrosis factor-α. Conclusions: Common structural and functional features between human and mouse NOD2 were identified. This should allow for development of relevant animal models to evaluate the role of NOD2 in chronic inflammatory disorders.

Original languageEnglish (US)
Pages (from-to)272-276
Number of pages5
JournalInflammation Research
Volume52
Issue number6
StatePublished - Jun 1 2003

Fingerprint

Cloning
Organism Cloning
NF-kappa B
Genes
Tissue
Chemical activation
Thymus
Tissue Distribution
Transcription
Chromosomes
Monocytes
Interferons
Distribution functions
Lipopolysaccharides
Exons
Assays
Skin
Animals
Kidney
Complementary DNA

Keywords

  • Blau Syndrome
  • CARD15 gene
  • Crohn's disease
  • NF-kappa B
  • NOD2 gene

ASJC Scopus subject areas

  • Toxicology
  • Pharmacology (medical)
  • Immunology
  • Cell Biology

Cite this

Cloning, sequencing and expression analysis of the mouse NOD2/CARD15 gene. / Iwanaga, Y.; Davey, Michael; Martin, Tammy; Planck, Stephen; DePriest, M. L.; Baugh, M. M.; Suing, C. M.; Rosenbaum, James (Jim).

In: Inflammation Research, Vol. 52, No. 6, 01.06.2003, p. 272-276.

Research output: Contribution to journalArticle

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