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Caspase-4 knockout in THP-1 cells

Inflammasome activation in THP1-KO-CASP4 cells

Human caspase-4 (CASP4) and CASP5, and their murine ortholog CASP11 are a family of inflammatory caspases that play a crucial role in the non-canonical inflammasome responses to intracellular lipopolysaccharide (LPS) [1-3].

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InvivoGen has generated THP1-KO-CASP4 cells from the human monocytic THP1 cell line THP1-Null2 through the stable knockout of the caspase-4 (CASP4) gene.

• THP1-KO-CASP4 cells – Knockout (KO) of the CASP4 gene

In this cell line, mature IL-1β secretion is not affected when the canonical inflammasome is activated. However, mature IL-1β secretion is severely impaired upon activation of the non-canonical inflammasome by intracellular LPS. The remnant activity may be attributed to a partial rescue of the non-canonical inflammasome response by CASP5, a functional homolog of CASP4. THP1-KO-CASP4 cells are useful to study the role of CASP4 in inflammasome responses and to screen novel therapeutics that target the CASP4 signaling pathways.

Features of THP1-KO-CASP4 cells:

Verified biallelic knockout of the CASP4 gene (both α and β isoforms) (DNA sequencing, PCR, and Western blot)
Severe impairment of mature IL-1β secretion upon non-canonical inflammasome activation only

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. Schmid-Burgk J.L. et al., 2015. Caspase-4 mediates non-canonical activation of the NLRP3 inflammasome in human myeloid cells. Eur. J. Immunol. 45:2911.
2. Baker P.J. et al., 2015. NLRP3 inflammasome activation downstream of cytoplasmic LPS recognition by both caspase-4 and caspase-5. Eur. J. Immunol. 45:2918.
3. Lagrange B. et al., 2018. Human caspase-4 detects tetra-acylated LPS and cytosolic Francisella and functions differently from murine caspase-11. Nat. Commun. 9:242.

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