The NBL1 Knockout Raji Polyclonal Cells consist of a heterogeneous population of Raji B lymphocytes engineered via CRISPR/Cas9-mediated disruption of the NBL1 gene. This polyclonal knockout pool provides a robust loss-of-function model for studying the functions of NBL1, a secreted DAN family bone morphogenetic protein (BMP) antagonist, in a Burkitt lymphoma background. The product is supplied as a viable, proliferating population suitable for immediate use in functional assays, bypassing the need for labor-intensive clone isolation. As a polyclonal knockout preparation, it captures the diversity of CRISPR-induced edits, enabling the assessment of gene disruption effects without clonal biases.
The Raji cell line is an EBV-positive B lymphoblastoid line originally derived from an 11-year-old male with Burkitt lymphoma. These cells exhibit features of germinal center B cells, express surface immunoglobulin, and retain active B cell receptor signaling pathways critical for lymphomagenesis. Raji cells are widely employed as a model system for studying B cell malignancies, immune evasion mechanisms, and oncogenic signaling networks. The presence of the EBV genome contributes to a partially activated state, enhancing the utility of these cells in dissecting the interplay between viral latency, cellular transformation, and tumor suppressor pathways.
NBL1 encodes a secreted BMP antagonist that directly binds BMP2, BMP4, and BMP7, preventing their interaction with type I receptors ACVR1 and BMPR1A/B. This inhibits SMAD1/5/8 phosphorylation and SMAD4-dependent transcription, attenuating expression of ID family inhibitors of differentiation (ID1, ID2, ID3). NBL1 signaling intersects with MAPK/ERK and Wnt pathways, and its expression is regulated by TP53, EGR1, and TGFB1. Other interacting factors include Gremlin and BMPER.
In the Raji B cell lymphoma context, NBL1 disruption enables dissection of BMP antagonism in lymphomagenesis. As a tumor suppressor, NBL1 loss may reveal altered proliferation, apoptosis, and differentiation driven by deregulated BMP signaling. The EBV-positive Raji line??s reliance on B cell receptor pathways provides a unique model to examine how loss of BMP inhibition impacts SMAD-dependent and non-canonical signaling, shedding light on the role of the BMP/TGF-?? axis in B cell transformation.
Researchers can utilize these polyclonal knockout cells for diverse assays, including phospho-SMAD1/5/8 Western blotting and ID1 RT-qPCR to monitor pathway activation, MTS/XTT proliferation assays, and Annexin V/7-AAD apoptosis analysis. Co-immunoprecipitation can verify disrupted NBL1-BMP2 interactions, while BRE-luciferase reporters quantify BMP pathway derepression. Additional applications include cell cycle analysis and xenograft tumor growth studies, supporting investigation of NBL1??s tumor-suppressive function in vivo. For technical inquiries, contact Ascent Research.
