The EYA3 Knockout Raji Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population in which the EYA3 locus has been targeted for gene disruption. Unlike monoclonal knockout lines, this polyclonal format preserves a spectrum of gene-edited alleles, allowing researchers to assess EYA3 function across a range of loss-of-function mutations. These cells serve as a powerful tool for investigating the role of EYA3 in transcriptional coactivation, tyrosine phosphatase activity, and oncogenic signaling in B cell lymphomas.
The parental Raji cell line, a human Burkitt lymphoma-derived lymphoblastoid B cell line, is EBV-positive and extensively employed to study B cell signaling, EBV biology, and lymphomagenesis. These cells exhibit high proliferation and deregulated MYC expression, mirroring aggressive lymphomas. The EYA3 knockout polyclonal cells generated on this background thus offer a valuable model for investigating the gene’s role in B cell malignancies.
EYA3 acts as a dual-function transcriptional coactivator and protein tyrosine phosphatase. It coactivates SIX1, SIX2, SIX4, and SIX5 transcription factors to induce CCND1 and MYC expression, driving cell proliferation. Upstream, YAP/TAZ-TEAD complexes promote EYA3 expression, linking Hippo signaling to cell cycle regulation. EYA3 also dephosphorylates H2AX and interacts with MDC1, facilitating DNA double-strand break repair via downstream mediators RBBP8 and CDC25A. It forms regulatory complexes with DACH1 to fine-tune target gene networks.
In Raji Burkitt lymphoma cells, EYA3 knockout disrupts SIX-dependent transcription of oncogenes, particularly CCND1 and MYC, leading to impaired G1/S transition and reduced proliferation. Concomitant loss of EYA3 phosphatase activity compromises DNA repair, as evidenced by persistent ??-H2AX foci and diminished MDC1 interaction, resulting in heightened apoptosis. Thus, this knockout model enables dissection of EYA3’s combined roles in proliferation and genome stability in a lymphomagenic context.
Researchers can confirm knockout by Western blotting, quantify SIX target gene expression by RT-qPCR, conduct proliferation and cell cycle analyses via flow cytometry, examine apoptosis, measure DNA repair by ??-H2AX foci formation, and perform xenograft tumor growth assays. This knockout population is ideal for investigating EYA3’s role in B cell lymphomagenesis and evaluating it as a therapeutic target in hematological malignancies. For additional information, please contact Ascent Research.
