Mouse ASC-expressing and Gasdermin D-KO RAW 264.7 Cells

ASC-OE & GSDMD-KO mouse macrophages

ABOUT

Gasdermin D knockout in RAW 264.7 cells

Gasdermin D (GSDMD) is a cytoplasmic protein with a pore-forming ability that has been described as a major actor in both canonical and non-canonical inflammasome responses [1, 2].

More details More details about GSDMD

 

To foster studies on GSDMD, InvivoGen has developed RAW-ASC KO-GSDMD cells, which were generated from the RAW-ASC cell line that derives from the naturally ASC deficient RAW 264.7 macrophage cell line [3]. RAW-ASC KO-GSDMD cells stably express the transfected murine ASC gene and have a stable knockout of the gasdermin D (GSDMD) gene.

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

In this cell line, both mature IL-1β secretion and pyroptotic cell death are abolished upon canonical and non-canonical inflammasome activation. This cell line is a useful tool to study the role of GSDMD in the inflammasome responses and is an alternative to the in vitro differentiation of mouse bone-marrow-derived macrophages. Additionally, it can be used as a control cell line for the screening of novel therapeutics that target GSDMD.

 

FEATURES OF RAW-ASC KO-GSDMD CELLS:

  • Verified biallelic knockout of the GSDMD gene and stable expression of the ASC gene (Western blot)
  • Complete abrogation of mature IL-1β secretion and pyroptosis after canonical and non-canonical inflammasome activation

 

Learn moreDownload our Practical guide on Inflammasomes

 

References:

1. Feng S. et al., 2018. Mechanisms of Gasdermin family members in inflammasome signaling and cell death. J. Mol. Biol. 430:3068.
2. Kovacs S.B. & Miao E.A. 2017. Gasdermins: effectors of pyroptosis. Trends Cell. Biol. 27:673.
3. 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.

Inflammasome signaling in RAW-ASC KO-GSDMD cells
Inflammasome signaling in RAW-ASC KO-GSDMD cells

SPECIFICATIONS

Specifications

Species
Murine
Tested applications

Complete abrogation of mature IL-1β secretion and pyroptosis after canonical and non-canonical inflammasome activation

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

Complete DMEM (see TDS)

Mycoplasma-free

Tested and validated using PlasmotestTM.

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    RAW-ASC KO-GSDMD Cells
  • Cat code: 
    raw-kogsdmd
  • 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

Details

GSDMD belongs to a family of six and ten gasdermins in humans and mice, respectively, which all have different expression patterns [1, 2]. GSDMD consists of two distinct domains, whereby the C-terminal domain exerts an auto-inhibitory function on the N-terminal domain.
GSDMD is cleaved by activated caspase-1 (CASP1) downstream of NLRP1, NLRP3, AIM2, NLRC4, or Pyrin canonical inflammasomes, or by activated CASP4/5 (human), CASP11 (mouse) non-canonical inflammasomes. The released GSDMD N-terminal domain oligomerizes to form 10-15 nm diameter pores at the cell membrane, thereby allowing the release of alarmins (e.g. HMGB1) and the secretion of mature IL-1β and IL-18 inflammatory cytokines. The accumulation of GSDMD pores in the membrane causes cell swelling and rupture, leading to an inflammatory cell death termed pyroptosis  [1, 2].
Importantly, GSDMD links the canonical and non-canonical inflammasome responses with the pore formation leading to stress signals, such as cytosolic ion concentration imbalances (i.e. K+ efflux) and ATP release. These signals induce the activation of NLRP3 and CASP1-mediated IL-1β/IL-18 maturation and secretion [3, 4].

 

1. Feng S. et al., 2018. Mechanisms of Gasdermin family members in inflammasome signaling and cell death. J. Mol. Biol. 430:3068.
2. Kovacs S.B. & Miao E.A. 2017. Gasdermins: effectors of pyroptosis. Trends Cell. Biol. 27:673.
3. Groslambert M. & Py B. 2018. Spotlight on the NLRP3 inflammasome pathway. J. Inflamm. Res. 11:359.
4. Mathur A. et al., 2017. Molecular mechanisms of inflammasome signaling. J. Leuk. Biol. 103:233.

DOCUMENTS

Documents

RAW-ASC KO-GSDMD Cells

Technical Data Sheet

Safety Data Sheet

Validation Data Sheet

Certificate of analysis

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