NLRP3 Knockout Cell Line (MARC145)

The choice depends on whether you are studying NLRP3 (NLR family pyrin domain containing 3, cryopyrin)'s role as the principal canonical inflammasome scaffold or modeling its applications in inflammatory disease and emerging viral immunology. The Knockout line is the standard tool for asking whether NLRP3 is required for these processes — NLRP3 is the most studied inflammasome sensor, activated by an extraordinarily broad range of danger signals (cholesterol crystals, monosodium urate crystals in gout, silica, asbestos, ATP, pore-forming toxins, ROS, mitochondrial damage, viral infections); upon activation, NLRP3 oligomerizes and recruits the adaptor ASC/PYCARD and pro-caspase-1, assembling the canonical NLRP3 inflammasome that drives IL-1β/IL-18 maturation and gasdermin D-mediated pyroptosis. Overexpression is useful for studying NLRP3 gain-of-function effects. For viral immunology and inflammasome research, the EDITGENE NLRP3 Knockout in MARC145 is uniquely valuable — MARC145 is the principal in vitro host for ⭐⭐ porcine reproductive and respiratory syndrome virus (PRRSV), one of the most economically important swine pathogens; PRRSV-induced inflammasome activation is an emerging research area. Rescue with wild-type, NACHT-domain-mutant (R260W gain-of-function CAPS mutation), or PYD-domain-mutant NLRP3 enables comprehensive structure-function and disease modeling. The knockout is a critical specificity tool for ⭐⭐ NLRP3 inhibitors: ⭐⭐ MCC950/CRID3 (the prototype selective NLRP3 inhibitor, basis for ⭐ inzomelid/IFM-2427 and dapansutrile/OLT1177 in clinical development for CAPS, gout, cardiovascular inflammation, and other inflammatory diseases), ⭐ colchicine (FDA-approved for gout and recently cardiovascular indications based in part on NLRP3 inhibition); for ⭐ ⭐ Cryopyrin-Associated Periodic Syndromes (CAPS) modeling; and for studying NLRP3 in atherosclerosis, gout, Alzheimer's disease, and PRRSV-induced inflammation.

Primary applications: • NLRP3 inflammasome activation: LPS+nigericin, LPS+ATP, monosodium urate crystal, or PRRSV-induced inflammasome activation (caspase-1 cleavage, IL-1β maturation, GSDMD-N pyroptosis) — abolished in NLRP3 KO. • ASC speck formation: ASC speck imaging analysis in NLRP3-null cells. • PRRSV-inflammasome interaction: in MARC145 PRRSV-permissive context, characterization of PRRSV-induced NLRP3 inflammasome activation. • NLRP3 inhibitor specificity: critical genetic control for ⭐⭐ MCC950/CRID3, ⭐ inzomelid (IFM-2427), ⭐ dapansutrile (OLT1177) — all in clinical development for CAPS/gout/cardiovascular inflammation. • CAPS modeling: rescue with patient-derived NACHT-domain CAPS mutations (R260W, etc.) for Cryopyrin-Associated Periodic Syndromes disease modeling. • Comparative immunology: cross-species NLRP3 inflammasome analysis given the African green monkey origin of MARC145 — useful for primate vs human vs porcine inflammasome comparison. EDITGENE recommends this MARC145-based model as a uniquely valuable tool for PRRSV-inflammasome research, cross-species inflammasome biology, and emerging NLRP3-targeted inflammatory disease drug development — bridging veterinary virology and human inflammation therapeutics.

Yes. NLRP3 rescue experiments are gold-standard for inflammasome research: • Construct design: use a codon-modified NLRP3 sequence with a small C-terminal tag (FLAG, HA). NLRP3 has N-terminal PYD (pyrin) domain (ASC binding), central NACHT/NOD ATPase domain (oligomerization), and C-terminal LRR (leucine-rich repeat) domain (autoinhibition/sensing) — preserve all elements. • CAPS gain-of-function rescue: ⭐ R260W, D305N, T348M, A352V NACHT-domain mutations are characteristic CAPS-causing variants conferring constitutive NLRP3 activation — gold-standard for CAPS disease modeling. • ATPase-dead rescue: Walker A/B motif mutations in the NACHT domain abolish oligomerization. • PYD-mutant rescue: PYD domain mutations disrupt ASC adaptor interaction. • Species considerations: African green monkey NLRP3 and human NLRP3 share extensive homology; human or species-matched cDNA can be used depending on experimental goals. • MCC950-resistant rescue: emerging characterized NLRP3 mutations conferring MCC950 resistance enable on-target inhibitor validation. • Functional readout: rescue should restore NLRP3 inflammasome activation, ASC speck formation, and pyroptosis in response to canonical activators (LPS+nigericin, MSU crystals). MARC145-specific considerations: • MARC145 is an African green monkey (Chlorocebus sabaeus) kidney epithelial cell line derived from MA-104 by selection for enhanced permissivity to porcine reproductive and respiratory syndrome virus (PRRSV) — MARC145 is the principal in vitro host for PRRSV propagation, isolation, and vaccine production. • Lentiviral transduction is supported with moderate efficiency; optimization may be required compared to standard human cell lines. • MARC145 cells retain CD163 expression (the principal PRRSV entry receptor) and Siglec-1/CD169 components of the PRRSV entry machinery. • As an African green monkey-derived line, MARC145 enables cross-species comparison of innate immune pathways (NLRP3, type I IFN, others) relative to human, porcine, and other mammalian systems.