The Ddr2 Knockout RAW 264.7 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout population of the murine RAW 264.7 macrophage-like cell line, featuring targeted disruption of the Ddr2 gene locus. This polyclonal loss-of-function model enables bulk functional studies without clonal isolation, offering a versatile tool for investigating Ddr2-dependent cellular processes. As a polyclonal pool, it minimizes clonal variation artifacts and is suited for assays requiring large cell numbers.
The host RAW 264.7 cell line, derived from a BALB/c mouse Abelson murine leukemia virus-induced tumor, is a widely employed monocyte/macrophage model. It exhibits characteristic phagocytic activity, robust inflammatory responses, and rapid proliferation, making it a standard system for macrophage biology, immune signaling, and host-pathogen interactions. Its established use in diverse in vitro assays ensures compatibility with existing experimental protocols.
Ddr2 encodes a receptor tyrosine kinase that is specifically activated by fibrillar collagens, including types I, II, and III. Upon collagen binding, Ddr2 undergoes autophosphorylation and recruits adaptor proteins such as Shc, Src, and FAK, initiating downstream cascades. Central pathways include Ras-Raf-MEK-ERK1/2 and PI3K-AKT signaling, along with JNK and p38 activation. These pathways converge on transcription factors like STAT3, NF-??B, and AP-1, which regulate expression of matrix metalloproteinases (MMP1, MMP2, MMP9), Snail, and other effectors controlling cell adhesion, migration, proliferation, and extracellular matrix remodeling.
In the macrophage context, Ddr2 signaling bridges collagen-rich matrix environments with immune cell function. RAW 264.7 macrophages respond to collagen via Ddr2 to modulate adhesion, migration, and MMP secretion, processes critical in tissue fibrosis, atherosclerotic plaque development, and chronic inflammation. Disruption of Ddr2 in these cells permits dissection of collagen-induced signaling events in the absence of confounding receptor activity, revealing the kinase??s contribution to macrophage polarization, phagocytic capacity, and cytokine production.
This knockout model supports a wide array of experimental applications. It is ideal for studying extracellular matrix signaling, macrophage-dependent fibrosis, tumor-associated macrophage function in cancer invasion, and inflammatory responses. Representative assays include collagen-stimulated phospho-ERK and phospho-AKT Western blotting, RT-qPCR for downstream targets, Transwell migration/invasion assays, adhesion to collagen substrates, flow cytometry for surface markers, and ELISA-based cytokine quantification. Additionally, it enables RNA-seq profiling and drug screening aimed at collagen receptor modulation. For further information or to request a quote, please contact Ascent Research.
