The MEGF8 Knockout Raji Polyclonal Cells are a polyclonal knockout cell population generated by CRISPR/Cas9-mediated disruption of the MEGF8 gene in the human Raji B lymphocyte cell line. This product provides a bulk-edited heterogeneous pool of MEGF8-deficient cells, enabling functional analysis of MEGF8 without clonal selection biases. The polyclonal format preserves population-level heterogeneity, suitable for studying gene function in a physiologically relevant context.
Raji cells are an immortalized human B lymphoblastoid cell line originally derived from a Burkitt’s lymphoma patient and are persistently infected with Epstein-Barr virus (EBV). They serve as a widely used model for B-cell biology, lymphomagenesis, and EBV-associated oncogenesis. The Raji line exhibits robust proliferation, well-characterized surface markers, and susceptibility to various signaling manipulations, making it a versatile platform for dissecting molecular pathways.
MEGF8 encodes a large transmembrane protein containing multiple EGF-like domains and functions as a receptor or co-receptor within the primary cilium. It modulates Hedgehog (HH) and bone morphogenetic protein (BMP) signal transduction by interacting with ligands such as Sonic hedgehog (SHH), Indian hedgehog (IHH), and BMP4, as well as receptors and ciliary complexes including the BBSome. Mechanistically, MEGF8 regulates downstream effectors: GLI transcription factors GLI1 and GLI2, PTCH1, and SMAD1/5/8 transcription factors. Loss of MEGF8 disrupts these signaling cascades, impairing GLI-mediated transcription and BMP-SMAD responses, which are essential for craniofacial morphogenesis and skeletal development. Representative pathway components include SHH, SMO, GLI1/2, PTCH1, BMP4, BMPR1A, and SMAD1/5/8.
While MEGF8 mutations are primarily linked to Carpenter syndrome and craniosynostosis, its knockout in Raji B lymphocytes offers a unique tool to explore HH and BMP signaling in lymphoid cells. Hedgehog and BMP pathways have emerging roles in B-cell proliferation, differentiation, and survival, and their dysregulation may intersect with EBV-driven lymphomagenesis. Thus, these knockout cells enable investigation of MEGF8-dependent signaling in an immune-cell context, extending beyond traditional developmental biology models.
These polyclonal knockout cells are suitable for a range of experimental applications, including functional studies of MEGF8 in HH/BMP signaling, craniosynostosis disease modeling, and screening for small-molecule modulators or genetic interactors. Representative assays include Western blot analysis for MEGF8 and downstream pathway proteins, RT-qPCR for HH and BMP target genes (e.g., GLI1, PTCH1, ID1), GLI-luciferase reporter assays to quantify pathway activity, immunofluorescence for ciliary protein localization, co-immunoprecipitation to map protein interactions, flow cytometry for surface marker profiling, and proliferation/apoptosis assays to assess phenotypic outcomes. The polyclonal nature allows assessment of knockout consequences in a mixed population, more closely mimicking in vivo heterogeneity. For additional information, please contact Ascent Research.
