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Murine NOD1 expressing HEK293 reporter cells

Signaling pathways in HEK-Blue™ mNOD1 cells

HEK-Blue™ mNOD1 cells were engineered from the human embryonic kidney HEK 293 cell line to assess the role of the Nucleotide-binding Oligomerization Domain-containing protein 1 (NOD1).

These cells stably express the murine (m) NOD1 gene and an NF-κB-inducible SEAP reporter gene. SEAP (secreted embryonic alkaline phosphatase) activity upon NOD1 stimulation can be readily determined by performing the assay in HEK-Blue™ Detection. This cell culture medium allows for real-time detection of SEAP. Alternatively, SEAP activity may be monitored using QUANTI-Blue™, a SEAP detection reagent.

The overexpression of mNOD1 renders HEK-Blue™ mNOD1 cells to strongly respond to NOD1-specific ligands (see figures). As HEK293 cells express endogenous levels of TLR3 and TLR5 [in-house data], HEK-Blue™ mNOD1 cells will respond to their cognate ligands such as Poly(I:C), and flagellin. They do not express human NOD2 (see figure).

NOD1 and NOD2 are cytosolic pattern recognition receptors (PRRs), specialized to sense distinct motifs of peptidoglycan, an essential constituent of the bacterial cell wall. NOD1 senses the D-γ-glutamyl-meso-DAP dipeptide (iE-DAP), which is found in PGN of all Gram-negative and certain Gram-positive bacteria [1].

More details

Key Features:

Verified overexpression of murine NOD1
Strong and stable response to NOD1-specific ligands
Distinct monitoring of NF-κB by assessing the SEAP activities

Applications:

Defining the distinct role of NOD1 in the NF-κB-dependent pathway
Screening for NOD1-specific agonists or inhibitors in comparison with their parental cell line HEK-Blue™ Null1

References:

1. Correa RG, Milutinovic S, Reed JC,. 2012. Roles of NOD1 (NLRC1) and NOD2 (NLRC2) in innate immunity and inflammatory diseases. Biosci Rep. 32(6):597-608.

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