Mouse ASC-expressing and NLRC4-KO RAW 264.7 Cells

ASC-OE & NLRC4-KO mouse macrophages

ABOUT

NLRC4 knockout in RAW 264.7 cells

NLRC4 (Nucleotide-binding domain (NBD) and leucin-rich repeat (LRR) receptor, CARD domain-containing protein 4, or IPAF) is a cytoplasmic protein which upon activation, triggers the assembly of an NLRC4 inflammasome [1-3].

More details More details about NLRC4

 

InvivoGen has generated RAW-ASC KO-NLRC4 cells from the RAW-ASC cell line, which derives from the naturally ASC-deficient RAW 264.7 macrophage cell line [4]. RAW-ASC KO-NLRC4 cells stably express the transfected murine ASC gene and feature a stable knockout of the NLRC4 gene. 

• RAW-ASC KO-NLRC4 cells – Knockout (KO) of the NLRC4 gene and expression of the murine ASC gene

These cells exhibit impaired IL-1β secretion and pyroptosis upon incubation with known inducers of the NLRC4 inflammasome. Importantly, IL-1β secretion and pyroptosis remain unaffected upon incubation with other inflammasome inducers such as Nigericin (NLRP3 inducer).

 

FEATURES OF RAW-ASC KO-NLRC4 CELLS:

  • Absence of functional NLRC4 protein expression
  • Verified biallelic knockout of the NLRC4 gene (DNA sequencing, PCR, and Western blot)
  • Altered IL-1β secretion and pyroptosis early upon NLRC4 inflammasome activation

 

Learn moreDownload our Practical guide on Inflammasomes

 

References:

1. Zhang L. et al., 2015. Cryo-EM structure of the activated NAIP2-NLRC4 inflammasome reveals nucleated polymerization. Science. 350:404-409.
2. Zhao Y. et al., 2011. The NLRC4 inflammasome receptors for bacterial flagellin and type III secretion apparatus. Nature. 477: 596-600.
3. Bauer R. & Rauch I., 2011. The NAIP/NLRC4 inflammasome in infection and pathology. Mol. Aspects Med. 2020 Jun 1:100863.
4. Pelerin P. et al., 2008. P2X7 receptor differentially couples to distinct release pathways for IL-1β in mouse macrophage. J. Immunol. 180:7147.

Disclaimer:  These cells are for internal research use only and are covered by a Limited Use License (See Terms and Conditions). Additional rights may be available.

SPECIFICATIONS

Specifications

Cell type
Macrophage
Growth properties
Adherent
Tissue origin
Mouse macrophages
Antibiotic resistance
Blasticidin
Growth medium

Complete DMEM (See TDS)

Mycoplasma-free

Verified using Plasmotest™

Quality control

Each lot is functionally tested and validated. 

Additional information

Test medium: DMEM without phenol red (See TDS). Note: Phenol red causes high background signal in the LDH (lactate dehydrogenase) assay used to monitor inflammasome-induced cell death.

CONTENTS

Contents

  • Product: 
    RAW-ASC KO-NLRC4 Cells
  • Cat code: 
    raw-konlrc4
  • Quantity: 
    3-7 x 10^6 cells
Includes:

1 ml of Normocin™ (50 mg/ml) 1 ml of Blasticidin (10 mg/ml)

Shipping & Storage

  • Shipping method:  Dry ice
  • Storage:

    • Liquid Nitrogen Vapor
    Stability: 20 passages

    Caution:

    • Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.

Details

Inflammasomes are cytoplasmic multi-protein complexes, characterized by a primary sensor, that assemble in response to infections and cellular damage. NLRC4 is an indirect sensor that must associate with NAIP (NLR family apoptosis inhibitory protein) to induce the assembly of an NLRC4 inflammasome. A single NAIP operates upstream of NLRC4 in humans and recognizes intracellular Flagellin, Needle, or Rod from several Gram-negative bacterial strains [1]. These ligands are sensed by NAIP upon bacterial invasion, or cytosolic translocation through the bacterial type III or IV secretion systems (T3SS or T4SS). Once recruited by NAIP, NLRC4 triggers a homo-polymerization through NBD-NBD interactions, allowing a CARD clustering [1]. The NLCR4 polymer further associates to pro-caspase-1, either through direct CARD-CARD interaction or through the binding of the ASC (apoptosis-associated speck-like protein) adaptor [1]. Activation of caspase-1 induces the maturation of pro-IL-1β/pro-IL-18, cleavage of the pore-forming gasdermin D (GSDMD), secretion of IL-1β/-18, and pyroptosis [1-3]. The NLRC4 inflammasome appears to protect mucosal barriers such as the lung, stomach, and intestine from invading bacteria [2]. Gain-of-function mutations have been described in human NLRC4 and are associated with auto-inflammatory conditions [3]. 

 

1. Zhang L. et al., 2015. Cryo-EM structure of the activated NAIP2-NLRC4 inflammasome reveals nucleated polymerization. Science. 350:404-409.
2. Zhao Y. et al., 2011. The NLRC4 inflammasome receptors for bacterial flagellin and type III secretion apparatus. Nature. 477: 596-600.
3. Bauer R. & Rauch I., 2011. The NAIP/NLRC4 inflammasome in infection and pathology. Mol. Aspects Med. 2020 Jun 1:100863.

DOCUMENTS

Documents

RAW-ASC KO-NLRC4 Cells

Technical Data Sheet

Validation Data Sheet

Safety Data Sheet

Certificate of analysis

Need a CoA ?

CUSTOMER SERVICE & TECHNICAL SUPPORT

Question about this product ?