The NFATC3 Knockout Raji Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population targeting the NFATC3 gene in the human Raji B lymphocyte line. These polyclonal cells are derived from the Raji host line through CRISPR/Cas9-mediated gene disruption, generating a heterogeneous pool of allelic modifications that collectively eliminate NFATC3 function. This product serves as a ready-to-use model for loss-of-function studies without requiring single-cell clone isolation, and it is supplied as a live cell stock suitable for direct experimental application or further manipulation.
The Raji host cell line was originally established from a Burkitt lymphoma patient and harbors the Epstein?CBarr virus (EBV) genome. These cells display characteristics of germinal center B cells and are widely used as a model system for B cell biology, lymphomagenesis, and EBV-driven oncogenesis. Raji cells grow in suspension and retain key B cell signaling pathways, including surface expression of the B cell receptor (BCR) and CD40, enabling investigation of proximal and distal signaling events relevant to immune activation and malignant transformation.
NFATC3 encodes a member of the nuclear factor of activated T cells (NFAT) transcription factor family that operates primarily downstream of calcium?Ccalcineurin signaling. In resting lymphocytes, NFATC3 is phosphorylated and retained in the cytoplasm. Upon receptor-mediated calcium mobilization??triggered through PLC?? activation and IP3 receptor-mediated store release??calcineurin binds calmodulin and dephosphorylates NFATC3, inducing its nuclear translocation. In the nucleus, NFATC3 cooperates with transcriptional partners such as AP-1 to drive expression of target genes including IL2, IL4, CD40LG, BCL2, and multiple cell cycle regulators. The protein also interacts with other NFAT isoforms (NFATC1, NFATC2), glycogen synthase kinase 3?? (GSK3B), and casein kinase 1 (CK1), which modulate its nuclear?Ccytoplasmic shuttling and transcriptional activity.
Disruption of NFATC3 in Raji cells provides a physiologically relevant model to dissect BCR- and CD40-driven calcium signaling and NFAT-dependent transcriptional programs in a B lymphoma background. Because NFATC3 is implicated in immune cell activation and proliferation, this knockout model is valuable for examining how calcineurin/NFAT pathways contribute to the transformed phenotype of EBV-positive B cells, and for exploring their role in B cell differentiation and survival. The polyclonal population captures diverse editing events, reflecting a more averaged biological response that can complement clonal studies.
Typical applications include Western blotting to verify NFATC3 protein loss, RT-qPCR to assess downstream target gene expression, flow cytometry for surface marker and activation profiling, and calcium flux assays to dissect proximal signaling. Proliferation and viability endpoints can be measured using MTT or CellTiter-Glo, and global transcriptional changes may be analyzed by RNA-seq. This model is well suited for drug screening targeting calcineurin or NFAT inhibitors, functional genomics screens, and studies of B cell receptor or cytokine receptor cross-talk. For further details, please contact Ascent Research.
