Sh2d3c Knockout RAW 264.7 Cell Line

The Sh2d3c Knockout RAW 264.7 Cell Line is a CRISPR/Cas9-edited knockout cell line that provides a targeted disruption of the Sh2d3c gene in a murine macrophage background. This model serves as a loss-of-function system for dissecting the roles of Sh2d3c in integrin-mediated adhesion, migration, and cytoskeletal reorganization. The knockout cell line is derived from the well-characterized RAW 264.7 macrophage line, enabling researchers to examine Sh2d3c-dependent signaling in an innate immune context.

RAW 264.7 is a murine macrophage cell line originally derived from BALB/c mice, widely adopted for studies of macrophage biology due to its functional responsiveness to lipopolysaccharide (LPS) and its capacity for phagocytosis, antigen presentation, and production of inflammatory cytokines such as TNF-?? and IL-6. These features make it a relevant model for investigating innate immune mechanisms, host-pathogen interactions, and inflammatory responses.

Sh2d3c encodes the adaptor protein NSP3/Chat, which serves as a molecular scaffold linking integrin and T cell receptor (TCR) signaling to the actin cytoskeleton. Following integrin ligation or TCR engagement, Src family kinases such as Lck and Fyn phosphorylate Sh2d3c, facilitating its interaction with p130Cas (BCAR1) and Crk. This assembly promotes the activation of Rac1, leading to actin polymerization, focal adhesion turnover, and enhanced cell migration. By coordinating these events, Sh2d3c operates downstream of integrin receptors, chemokine receptors, and FAK to regulate adhesion and motility.

In macrophages, Sh2d3c-mediated signaling is implicated in the regulation of integrin-dependent adhesion, migration, and phagocytosis??processes essential for tissue surveillance, wound healing, and immune effector functions. Disruption of Sh2d3c in the RAW 264.7 background allows for the direct assessment of its contribution to macrophage motility, phagocytic uptake, and integrin signaling dynamics. This knockout model is therefore valuable for studying inflammatory disorders, autoimmune diseases, and tumor-associated macrophage behavior relevant to cancer metastasis.

This cell line supports a range of functional assays to interrogate Sh2d3c biology, including Transwell migration assays, adhesion assays on extracellular matrix substrates, and phagocytosis assays using fluorescent particles or bacteria. Downstream signaling can be examined by western blotting for phosphorylated FAK and p130Cas, while immunofluorescence microscopy reveals actin cytoskeletal organization. Flow cytometry enables quantification of integrin surface expression, and co-immunoprecipitation can confirm disrupted Sh2d3c-p130Cas complexes. The Sh2d3c Knockout RAW 264.7 Cell Line thus provides a robust platform for dissecting macrophage integrin signaling pathways with potential translational implications in immune dysfunction and cancer. For further information, please contact Ascent Research.