The FZD6 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population engineered to disrupt the FZD6 gene in human Raji B lymphocytes. This product provides a loss-of-function model for investigating the role of Frizzled-6, a receptor in non-canonical Wnt signaling, within a Burkitt??s lymphoma-derived background. The polyclonal nature of the knockout pool captures diverse editing events, enabling robust functional studies without clonal isolation.
The Raji cell line, established from a Burkitt??s lymphoma patient, is an EBV-positive, lymphoblastoid B-lymphocyte model widely utilized for studying B-cell antigen presentation, immunoglobulin secretion, and EBV-mediated oncogenesis. Its transformed phenotype retains many features of mature B cells, making it an ideal platform for interrogating signal transduction pathways in lymphoma.
FZD6 encodes Frizzled-6, a seven-transmembrane receptor that transduces non-canonical Wnt/planar cell polarity (PCP) signals upon binding ligands such as WNT5A, WNT4, and WNT9B. Upon activation, FZD6 recruits Dishevelled (DVL1/2/3), engaging core PCP proteins VANGL1/2 and CELSR1?C3. This cascade activates small GTPases RAC1 and RHOA, leading to JNK and c-Jun phosphorylation and subsequent AP-1 transcriptional regulation of genes controlling cytoskeletal dynamics and cell migration. Additional interacting factors include GNAQ/GNA11, ARRB1, and PKC, while downstream effectors such as CDC42 and PAK1 orchestrate cytoskeletal rearrangements. Notably, FZD6 signaling can antagonize canonical Wnt/??-catenin pathway output.
In Raji B cells, FZD6 knockout disrupts non-canonical Wnt/PCP signaling, enabling dissection of its roles in cell polarity, directed migration, and invasive behavior. Aberrant Wnt signaling is implicated in B-cell lymphoma progression and metastasis, and FZD6 itself has been associated with cancer cell dissemination in melanoma, breast, and gastric carcinomas, as well as with congenital nail clubbing. Thus, this knockout model allows investigation of FZD6-dependent mechanisms underlying malignant B-cell phenotypes.
Researchers can employ these polyclonal knockout cells in a broad array of functional assays. Western blotting for FZD6 and phospho-JNK confirms target disruption and pathway inactivation, while transwell migration assays quantify changes in chemotactic motility. Cell viability (MTT) and luciferase reporter assays (TOP/FOP for ??-catenin, AP-1 for non-canonical output) measure transcriptional activity. Immunofluorescence, co-immunoprecipitation with DVL, and flow cytometry for surface FZD6 enable analysis of subcellular localization and protein interactions, and RNA-seq can profile downstream transcriptional changes. This model is suitable for drug target validation and functional dissection of Wnt/PCP signaling in B-cell lymphoma. For further information, please contact Ascent Research.
