The MDFIC Knockout Raji Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Raji B lymphocyte line, engineered to disrupt the transcriptional repressor MDFIC. This pooled knockout model enables loss-of-function analysis of MDFIC in a lymphoid background, circumventing clonal selection artifacts. The polyclonal format preserves heterogeneous editing outcomes, providing a robust population-level functional assessment while maintaining the parental line??s suspension growth properties. Researchers can employ this system to interrogate MDFIC-dependent gene regulation without the constraints of single-cell-derived genotypes, offering a versatile platform for Wnt and TCF/LEF pathway studies.
The host Raji cell line originates from an EBV-positive Burkitt lymphoma and exhibits characteristic B lymphocyte features, including surface immunoglobulin expression and MHC class II-mediated antigen presentation. Widely adopted in immunology and oncology, Raji cells serve as a model for B-cell malignancies, antibody production, and immune surveillance mechanisms. Their Epstein?CBarr virus latency III program confers constitutive NF-??B and STAT signaling, rendering them sensitive to microenvironmental cues. Suspension-adapted growth in RPMI-1640 with serum facilitates scalable culture for assays demanding viable lymphoid cells, such as functional apoptosis screens or drug sensitivity profiling.
MDFIC encodes a transcriptional repressor that antagonizes Wnt/??-catenin signaling by competing with ??-catenin for binding to TCF/LEF transcription factors, notably TCF7L2. In the nucleus, MDFIC displaces ??-catenin from TCF/LEF complexes, thereby inhibiting expression of Wnt target genes including MYC, CCND1, and AXIN2. Upstream Wnt ligands such as Wnt3a and Wnt1 engage Frizzled-LRP6 receptor complexes to stabilize ??-catenin, which normally associates with TCF7L2 to drive proliferation. MDFIC also represses MyoD-mediated myogenic transcription and interacts with p32/C1QBP and HIV-1 Tat, linking it to viral transactivation. Knockout of MDFIC lifts this tonic repression, resulting in heightened TCF/LEF-dependent transcription and potentially altering B-cell growth programs.
In Raji B lymphocytes, MDFIC loss disrupts the negative feedback loop that modulates Wnt-dependent proliferation and survival. Constitutively active Wnt signaling can drive MYC and CCND1 upregulation, mimicking features of aggressive lymphomas and primary immunodeficiencies associated with MDFIC variants. Because Raji cells are also a model for HIV-1 Tat-mediated transcription, MDFIC knockout permits examination of how Tat hijacks host repressors to promote viral gene expression. This model thus bridges lymphomagenesis, immunodeficiency, and host?Cpathogen interactions, enabling dissection of MDFIC??s tumor-suppressive roles in the context of EBV-driven B-cell transformation.
Applications of this polyclonal knockout population span functional genomics, signaling pathway reconstruction, and drug discovery. Users can quantify transcriptomic changes via RNA-seq or RT-qPCR for MYC and CCND1, validate protein-level effects through Western blotting of MDFIC and Wnt targets, and assess ??-catenin phosphorylation states. Flow cytometry enables profiling of B-cell markers and apoptosis, while TCF/LEF luciferase reporter assays directly measure Wnt transcriptional output. Drug sensitivity studies with Wnt inhibitors (e.g., porcupine or tankyrase inhibitors) can evaluate target dependency, and HIV-1 Tat co-expression experiments may elucidate MDFIC??s role in viral replication. For further details, please contact Ascent Research.
