The Piezo1 Knockout RAW 264.7 Cell Line is a CRISPR/Cas9-edited murine macrophage cell line with targeted disruption of the Piezo1 gene, providing a loss-of-function model for studying mechanotransduction. Derived from the RAW 264.7 line, it allows dissection of Piezo1-dependent signaling without variable endogenous expression, supporting research in macrophage biology including inflammation, phagocytosis, and migration.
RAW 264.7 is a well-characterized macrophage line established from a BALB/c mouse ascites tumor, widely used for studies of immune function. These cells display robust phagocytic activity, respond to LPS by secreting cytokines like TNF-?? and IL-6, and serve as a standard model for NF-??B and MAPK/ERK signaling. The Piezo1 knockout retains these properties while enabling focused analysis of mechanical force transduction.
PIEZO1 is a mechanosensitive cation channel that opens in response to membrane stretch, shear stress, or chemical agonist Yoda1, allowing calcium influx that activates calpain, calmodulin-dependent kinases such as CAMKII, and transcription factors NF-??B p65 and AP-1. This channel also interacts with STIM1 and the actin cytoskeleton, and couples to integrin-mediated adhesion and Rho GTPases to regulate cytoskeletal dynamics. In immune contexts, PIEZO1 signaling may intersect with TLR4-MyD88 pathways, linking mechanical and inflammatory stimuli.
In macrophages, PIEZO1 translates physical cues from the microenvironment into calcium-driven functional responses, potentially modulating phagocytosis, cytokine production, and migration. This knockout line enables specific testing of whether PIEZO1-mediated mechanotransduction is required for NF-??B-dependent inflammatory gene expression or phagocytic uptake. By decoupling mechanical input from calcium signaling, researchers can dissect the pathway logic governing macrophage activation in health and disease.
Typical applications include calcium imaging (Fluo-4) to compare mechanical or Yoda1-evoked transients, patch clamp recording of mechanosensitive currents, and Western blotting for phosphorylated ERK and p65. ELISA quantifies secreted TNF-?? and IL-6 under combined stimuli, and phagocytosis or migration assays reveal functional contributions of PIEZO1. This cell line also supports drug screening for Piezo1 modulators and studies of macrophage behavior in stiff tumor microenvironments. Contact Ascent Research for further technical details.
