ENG Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the ENG gene in the human Raji B lymphoblast cell line. This product provides a loss-of-function model for the transmembrane glycoprotein endoglin (CD105), a co-receptor of the transforming growth factor-beta (TGF-??) superfamily. The polyclonal format ensures a heterogeneous pool of edited cells, enabling robust population-level analyses without clonal artifacts. Researchers can employ this tool to systematically dissect endoglin-dependent signaling cascades and cellular responses in a lymphoid context relevant to immunology and oncology.
The host Raji cell line was originally established from a Nigerian patient with Burkitt lymphoma and is Epstein-Barr virus (EBV)-positive, exhibiting a mature B-cell phenotype and high transfection efficiency. These characteristics make Raji cells a widely accepted model for B-cell lymphoma biology and immune system studies, particularly for examining viral oncogenesis and lymphoproliferative mechanisms. The cells?? EBV status and rapid proliferation render them invaluable for investigating tumor microenvironment interactions and therapeutic vulnerabilities in aggressive B-cell malignancies.
Endoglin functions as a co-receptor that binds TGF-??1, TGF-??3, and bone morphogenetic protein 9 (BMP9) to modulate downstream Smad-dependent signaling. In endothelial cells, endoglin preferentially facilitates ALK1-mediated phosphorylation of Smad1/5/8, whereas its role in B cells may involve ALK5/Smad2/3 cascades. Upstream regulators include TGF-??1, HIF-1??, and VEGF, while downstream targets encompass Smad1/5/8, Smad2/3, and transcriptional effectors such as ID1. Interacting partners like TGF-?? receptor type II (T??RII), ALK1, ALK5, and integrins further contextualize its integration into broader angiogenic and immune-regulatory networks.
In the Raji host background, ENG knockout disrupts endoglin??s capacity to coordinate TGF-?? responsiveness, providing a precise tool to evaluate its contributions to B-lymphoma cell proliferation, survival, apoptosis, and immune evasion. This model permits dissection of TGF-??-mediated cross-talk with oncogenic pathways driven by EBV, offering insights into how endoglin may influence B-cell activation, differentiation, and cytokine secretion within the tumor microenvironment. Comparative studies between knockout and wild-type Raji cells can reveal endoglin??s modulatory effects on signaling thresholds that govern malignant B-cell behavior.
Typical applications include flow cytometry for CD105 surface expression, western blotting of phosphorylated Smad1/5/8 and Smad2/3, RT-qPCR quantification of TGF-?? target genes (e.g., ID1, SERPINE1), and proliferation or apoptosis assays under TGF-??1 stimulation. The knockout cells are suitable for co-immunoprecipitation of receptor complexes, immunofluorescence localization studies, and RNA-seq transcriptomic profiling to map endoglin-dependent gene networks. This model supports drug target validation for anti-angiogenic and TGF-?? pathway inhibitors, as well as functional screens for modulators of the endoglin/TGF-?? axis in B-cell contexts. For further information, please contact Ascent Research.
