THP1-KO-GSDMD Cells
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Cat.code:
thp-kogsdmdz
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ABOUT
Gasdermin D knockout in THP-1 cells
Gasdermin D (GSDMD) is a cytoplasmic protein with a pore-forming ability that has been described as a major actor in early canonical and non-canonical inflammasome responses [1]. GSDMD pores are formed via its N-terminal domain following cleavage of the full-length GSDMD protein by caspase-1 or human CASP4/5 (CASP11 in mice). These pores allow the release of alarmins (e.g. HMGB1) and the secretion of mature IL-1β and IL-18 inflammatory cytokines.
InvivoGen has developed THP1-KO-GSDMD cells, which were generated from the human monocytic THP-1 cell line THP1-Null2, through the stable knockout of the GSDMD gene.
• THP1-KO-GSDMD cells – Knockout (KO) of the GSDMD gene
These cells exhibit impaired early IL-1β secretion and pyroptosis responses upon canonical and non-canonical inflammasome activation. Although these cells are fully KO for the GSDMD protein, mature IL-1β secretion is detected upon prolonged incubation with some inflammasome inducers. This observation illustrates the reported IL-1β secretion upon GSDMD-independent cell death [2, 3].
Features of THP1-KO-GSDMD cells:
- Generated from the parental cell line THP1-Null2
- Verified biallelic knockout of the GSDMD gene (DNA sequencing, PCR, and Western blot)
- Altered IL-1β secretion and pyroptosis early upon canonical and non-canonical inflammasome activation
For detecting and quantifying the release of mature human (h)IL-1β, InvivoGen provides HEK-Blue™ IL-1β sensor cells, which express an NF-κB-inducible SEAP reporter gene. QUANTI-Blue™ Solution allows rapid colorimetric detection and measure of SEAP activity by reading the optical density at 630-650 nm.
Download our Practical guide on Inflammasomes
References:
1. Kovacs S.B. & Miao E.A. 2017. Gasdermins: effectors of pyroptosis. Trends Cell. Biol. 27:673.
2. Schneider K.S. et al.,2018. The Inflammasome Drives GSDMD-Independent Secondary Pyroptosis and IL-1 Release in the Absence of Caspase-1 Protease Activity. Cell Rep. 21:3846.
3. Zeng C-H. et al., 2019. ATP induces caspase-3/gasdermin E-mediated pyroptosis in NLRP3 pathway-blocked murine macrophages. Apoptosis. 24(9-10):703-717.
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
Inflammasome activation
Complete RPMI 1640 (see TDS)
Verified using PlasmotestTM
Each lot is functionally tested and validated.
CONTENTS
Contents
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Product:THP1-KO-GSDMD Cells
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Cat code:thp-kogsdmdz
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Quantity:3-7 x 10^6 cells
- 1 ml of Zeocin® (100 mg/ml).
- 1 ml of Normocin™ (50 mg/ml).
Shipping & Storage
- Shipping method: Dry ice
- Liquid nitrogen vapor
- Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.
Storage:
Caution:
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, 3].
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 [2, 4].
1. Kovacs S.B. & Miao E.A. 2017. Gasdermins: effectors of pyroptosis. Trends Cell. Biol. 27:673.
2. Groslambert M. & Py B. 2018. Spotlight on the NLRP3 inflammasome pathway. J. Inflamm. Res. 11:359.
3. Feng S. et al., 2018. Mechanisms of Gasdermin family members in inflammasome signaling and cell death. J. Mol. Biol. 430:3068.
4. Mathur A. et al., 2017. Molecular mechanisms of inflammasome signaling. J. Leuk. Biol. 103:233.
DOCUMENTS
Documents
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
Need a CoA ?
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