The Lacc1 Knockout THP-1 Cell Line is a CRISPR/Cas9-edited knockout cell line generated from the human THP-1 acute monocytic leukemia cell line. This product features targeted disruption of the Lacc1 gene, creating a loss-of-function model that enables rigorous investigation of LACC1-dependent signaling and metabolic processes. Engineered via CRISPR/Cas9 genome editing, the cell line maintains the parental line??s capacity for suspension growth and phorbol ester-induced differentiation into macrophage-like cells, while providing a stable genetic background for mechanistic studies.
THP-1 is a widely employed human monocytic leukemia cell line that serves as a versatile model for monocyte and macrophage biology. In its undifferentiated state, THP-1 cells grow as a suspension culture, but treatment with phorbol 12-myristate 13-acetate (PMA) induces adherence, morphological spreading, and expression of macrophage markers. Differentiated THP-1 macrophages respond robustly to inflammatory stimuli such as lipopolysaccharide (LPS) and tumor necrosis factor-alpha (TNF-alpha), activating pathways that lead to NLRP3 inflammasome assembly and cytokine secretion. This property makes the cell line particularly suitable for examining the interplay between innate immune signaling and cellular metabolism.
LACC1 (laccase domain-containing 1) is a regulator of fatty acid oxidation and mitochondrial homeostasis that directly influences NLRP3 inflammasome activation and autophagy. It acts downstream of pattern recognition receptors and cytokines, controlling mitochondrial reactive oxygen species (ROS) production and caspase-1-mediated cleavage of pro-interleukin-1beta (IL-1beta) and pro-IL-18. LACC1 interacts with NOD2, RIPK2, and autophagy machinery components ATG5 and LC3. Key pathway elements include LPS/TLR4/NF-??B signaling, the NLRP3 inflammasome, and the fatty acid oxidation enzyme CPT1A. Disruption of LACC1 uncouples lipid metabolism from inflammasome activation, leading to dysregulated IL-1beta secretion and altered autophagic flux.
Within the THP-1 background, LACC1 knockout provides a physiologically relevant platform to dissect how mitochondrial metabolism governs innate immune responses in monocytic cells. Upon PMA differentiation, knockout macrophages display defects in mitochondrial oxidative capacity and autophagy, which directly impinge on NLRP3 inflammasome assembly and downstream cytokine release. This phenotype recapitulates key aspects of autoinflammatory disorders such as juvenile idiopathic arthritis, inflammatory bowel disease, and systemic lupus erythematosus, where LACC1 variants have been identified. Consequently, the cell line serves as a targeted tool for probing the molecular hierarchy from fatty acid oxidation to inflammatory pathology.
The Lacc1 Knockout THP-1 Cell Line is suited for a wide array of functional assays, including Western blotting and ELISA for NLRP3, caspase-1, and IL-1beta quantification, Seahorse metabolic flux analysis to assess mitochondrial respiration, and fluorescence-based detection of mitochondrial ROS. Immunofluorescence for LC3 puncta and RT-qPCR for inflammatory cytokine transcripts further enable autophagy and gene expression profiling. Additionally, the cell line can be utilized in phagocytosis assays and high-throughput drug sensitivity screens targeting LACC1-dependent pathways. For further details or to discuss custom applications, please contact Ascent Research.
