The PPARG Knockout THP-1 Cell Line is a genetically engineered human monocytic cell line in which CRISPR/Cas9-mediated gene editing has been employed to disrupt the PPARG locus, creating a functional knockout. This product provides a defined loss-of-function system for investigating PPARG-dependent regulatory mechanisms in a monocyte/macrophage context. The cell line is offered as a live culture, amenable to standard tissue culture protocols and differentiation into macrophage-like cells using phorbol 12-myristate 13-acetate (PMA).
THP-1 is a spontaneously immortalized human monocytic cell line originally isolated from the peripheral blood of a one-year-old male with acute monocytic leukemia. It serves as a widely accepted model for studying monocyte-to-macrophage differentiation, innate immune responses, and inflammatory signaling. Upon PMA stimulation, THP-1 cells undergo growth arrest, become adherent, and develop characteristics of mature macrophages, including phagocytic activity, upregulation of surface antigens such as CD11b and CD36, and secretion of cytokines.
PPARG (peroxisome proliferator-activated receptor gamma) encodes a ligand-activated transcription factor belonging to the nuclear receptor superfamily. It forms obligate heterodimers with retinoid X receptor (RXR) and occupies PPAR response elements (PPREs) in target gene regulatory regions. PPARG is activated by endogenous ligands including fatty acids and 15-deoxy-??12,14-prostaglandin J2, and by synthetic agonists such as thiazolidinediones. Its transcriptional activity is modulated by phosphorylation from MAP kinases and AMPK, and by recruitment of coactivators (PGC-1??, SRC-1) or corepressors (NCoR, SMRT). In macrophages, PPARG induces expression of lipid metabolism genes (CD36, FABP4, LPL) and adiponectin, while transrepressing pro-inflammatory cytokines such as TNF-?? and IL-6, thereby steering cells toward an anti-inflammatory M2-like state and maintaining lipid homeostasis.
Within the THP-1 macrophage model, PPARG is a key node linking metabolic sensing and inflammatory output. The PPARG knockout cell line enables researchers to dissect the specific contribution of this nuclear receptor to processes such as macrophage differentiation, foam cell formation, and cytokine production. The model is particularly informative for studying diseases where PPARG-mediated anti-inflammatory and lipid-handling functions are compromised, including atherosclerosis, non-alcoholic fatty liver disease, obesity, and type 2 diabetes. Direct comparison of wild-type and PPARG-null THP-1 macrophages permits attribution of phenotypes to PPARG-dependent signaling.
This knockout cell line supports a wide range of investigative techniques. Gene expression and chromatin-binding studies can employ RT-qPCR and ChIP-qPCR to profile PPARG target genes and PPRE occupancy. Drug discovery applications include screening PPARG agonists or antagonists using luciferase reporter assays and assessing functional lipid uptake via oxidized LDL. Cytokine ELISA for TNF-?? and IL-10, along with flow cytometry for M1/M2 surface markers, characterize inflammatory responses and polarization. Western blotting confirms ablation of PPARG protein. This cell line is a precise genetic tool for metabolic inflammation research and preclinical evaluation of PPARG modulators. For inquiries, contact Ascent Research.
