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

Y. Iwanaga; M. P. Davey; T. M. Martin; S. R. Planck; M. L. DePriest; M. M. Baugh; C. M. Suing; J. T. Rosenbaum

doi:10.1007/s00011-003-1170-z

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

Y. Iwanaga, M. P. Davey, T. M. Martin, S. R. Planck, M. L. DePriest, M. M. Baugh, C. M. Suing, J. T. Rosenbaum

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40 Scopus citations

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 language	English (US)
Pages (from-to)	272-276
Number of pages	5
Journal	Inflammation Research
Volume	52
Issue number	6
DOIs	https://doi.org/10.1007/s00011-003-1170-z
State	Published - Jun 1 2003

Keywords

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

ASJC Scopus subject areas

Immunology
Pharmacology

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10.1007/s00011-003-1170-z

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@article{6dc644a4b9894aa88c39b1661fa815f9,

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

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.",

keywords = "Blau Syndrome, CARD15 gene, Crohn's disease, NF-kappa B, NOD2 gene",

author = "Y. Iwanaga and Davey, {M. P.} and Martin, {T. M.} and Planck, {S. R.} and DePriest, {M. L.} and Baugh, {M. M.} and Suing, {C. M.} and Rosenbaum, {J. T.}",

note = "Funding Information: Acknowledgments. The authors would like to thank Sunwen Chou (Portland VAMC) for access to the ABI PRISM 7700 unit, Anne Senters for DNA sequencing, and Robert Klein and Amy Carlos for northern blot analysis. This work was supported by NIH grants EY06484, EY10572 and EY13139; Research to Prevent Blindness awards to JTR, SRP and the Casey Eye Institute; the Rosenfeld family trust and the Department of Veterans Affairs.",

year = "2003",

month = jun,

day = "1",

doi = "10.1007/s00011-003-1170-z",

language = "English (US)",

volume = "52",

pages = "272--276",

journal = "Inflammation Research",

issn = "1023-3830",

publisher = "Birkhauser Verlag Basel",

number = "6",

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TY - JOUR

T1 - Cloning, sequencing and expression analysis of the mouse NOD2/CARD15 gene

AU - Iwanaga, Y.

AU - Davey, M. P.

AU - Martin, T. M.

AU - Planck, S. R.

AU - DePriest, M. L.

AU - Baugh, M. M.

AU - Suing, C. M.

AU - Rosenbaum, J. T.

N1 - Funding Information: Acknowledgments. The authors would like to thank Sunwen Chou (Portland VAMC) for access to the ABI PRISM 7700 unit, Anne Senters for DNA sequencing, and Robert Klein and Amy Carlos for northern blot analysis. This work was supported by NIH grants EY06484, EY10572 and EY13139; Research to Prevent Blindness awards to JTR, SRP and the Casey Eye Institute; the Rosenfeld family trust and the Department of Veterans Affairs.

PY - 2003/6/1

Y1 - 2003/6/1

N2 - 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.

AB - 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.

KW - Blau Syndrome

KW - CARD15 gene

KW - Crohn's disease

KW - NF-kappa B

KW - NOD2 gene

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U2 - 10.1007/s00011-003-1170-z

DO - 10.1007/s00011-003-1170-z

M3 - Article

C2 - 12835899

AN - SCOPUS:0037709366

SN - 1023-3830

VL - 52

SP - 272

EP - 276

JO - Inflammation Research

JF - Inflammation Research

IS - 6

ER -

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

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Keywords

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