The NLRP3 Knockout Marc-145 Cell Line is a CRISPR/Cas9-edited knockout model derived from the Chlorocebus aethiops (African green monkey) kidney epithelial Marc-145 cell line. This cell line features targeted disruption of the NLRP3 gene, providing a stable loss-of-function system for investigating NLRP3-mediated inflammasome signaling and innate immunity.
Marc-145 cells originate from the MA-104 African green monkey kidney epithelial line and display adherent growth. They are widely recognized for their permissiveness to viral replication, especially Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), and serve as an important model for viral infectivity and host innate immune responses in kidney-derived cells.
NLRP3 functions as a cytosolic sensor for diverse danger signals, including ATP (via P2RX7), K+ efflux, reactive oxygen species, and lysosomal damage. Upon activation, NLRP3 assembles the inflammasome complex through interactions with ASC (PYCARD) and NEK7, triggering pro-caspase-1 (CASP1) autoactivation. Active CASP1 processes pro-IL-1?? and pro-IL-18 into mature cytokines and cleaves gasdermin D (GSDMD) to induce pyroptosis, releasing HMGB1 and other inflammatory mediators. Priming of NLRP3 expression occurs through NF-??B signaling downstream of Toll-like receptors (e.g., TLR4) in response to microbial PAMPs. Key pathway components include NLRP3, PYCARD, CASP1, IL1B, IL18, GSDMD, P2RX7, TLR4, NFKB, and NEK7.
In Marc-145 cells, NLRP3 knockout enables dissection of inflammasome-dependent responses to viral infection and epithelial injury. Given the cell line??s susceptibility to PRRSV and its renal epithelial background, this model is particularly valuable for elucidating how NLRP3 influences cytokine production, pyroptotic cell death, and viral replication dynamics in kidney-derived innate immunity contexts.
Typical research applications include inflammasome mechanism studies, screening of NLRP3 inhibitors, and viral immunology investigations. Commonly employed assays are Western blotting for IL-1?? and caspase-1 processing, ELISA for released IL-1??/IL-18, LDH release pyroptosis assays, ASC speck visualization by immunofluorescence, RT-qPCR for inflammasome genes, and virus infectivity measurements. Co-immunoprecipitation can be used to probe NLRP3-ASC interactions. For further details, contact Ascent Research.
