The NR4A1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the gene encoding NR4A1 (Nur77, NGFI-B), an orphan nuclear receptor and immediate-early transcription factor. This polyclonal knockout model provides a heterogeneous pool of Raji cells carrying diverse loss-of-function events at the NR4A1 locus, enabling robust population-level studies of NR4A1-dependent biology without clonal selection bias. The polyclonal format is particularly suitable for functional screens and pooled analyses where representation of multiple genetic lesions is advantageous. As a ready-to-use reagent, these cells are intended for direct application in downstream assays following thawing and expansion, reducing the timeline for generating knockout models in B-lymphoblastoid cells.
Derived from a patient with Burkitt lymphoma, the Raji cell line is an Epstein-Barr virus (EBV)-positive B lymphoblastoid model that has been instrumental in immunology and cancer research for decades. Raji cells retain key features of mature B lymphocytes, including surface immunoglobulin expression and the capacity for antigen presentation. Their robust growth in suspension culture and well-characterized signaling networks make them an ideal host for studying B cell receptor (BCR) signaling, apoptosis regulation, and oncogenic mechanisms. The EBV-positive background further allows investigation of viral interactions with host cellular pathways, including NF-??B and survival signaling, which are often dysregulated in lymphomagenesis.
NR4A1 functions as a dual-action stress sensor that can either promote or suppress cell survival depending on the cellular context and post-translational modifications. In B cells, NR4A1 is rapidly induced by BCR stimulation and downstream kinases such as SYK, BTK, and PKC, acting through transcription factors like NF-??B, CREB, and MEF2. Its transcriptional targets include pro-apoptotic genes BIM (BCL2L11), FAS ligand, and TRAIL, as well as negative regulators of the cell cycle like p21/CDKN1A. Additionally, NR4A1 can translocate to mitochondria in response to apoptotic signals, where it physically interacts with BCL2, exposing the BCL2 BH3 domain and inducing cytochrome c release, caspase-9 and caspase-3 activation, and intrinsic apoptosis. NR4A1 also interacts with cofactors such as RXR, p300/CBP, and PARP1 to modulate inflammatory cytokines like IL-6 and metabolic targets including GLUT4, linking its activity to broader inflammatory and metabolic pathways.
In the Raji B-lymphoblastoid context, disrupting NR4A1 provides a powerful tool to dissect the balance between BCR-driven proliferation and apoptosis. Given the cell line’s origin from Burkitt lymphoma, a malignancy characterized by MYC translocations and aberrant survival signaling, NR4A1 knockout Raji cells enable researchers to examine how loss of this pro-apoptotic factor contributes to chemoresistance and oncogenic transformation. The model is particularly relevant for probing crosstalk between the NF-??B pathway, which is constitutively active in many B-cell lymphomas, and NR4A1-mediated death signals. By ablating NR4A1, users can interrogate its role in modulating BCL2 family members, mitochondrial integrity, and cytokine production, providing insights into therapeutic strategies that target lymphomas resistant to conventional treatments.
Typical research applications include elucidating NR4A1-dependent mechanisms in B cell lymphoma, studying BCR signal transduction kinetics, and investigating apoptosis and drug resistance. These cells can be used in Western blotting, annexin V-based apoptosis assays, caspase activity measurements, RT-qPCR for downstream gene expression, and flow cytometry to assess mitochondrial membrane potential and cell cycle distribution. Co-immunoprecipitation assays enable analysis of NR4A1-BCL2 interactions, while NF-??B luciferase reporters and phospho-signaling analysis facilitate pathway interrogation. Additional applications encompass screening for small-molecule modulators of NR4A1 and metabolic reprogramming studies in cancer. For further information, please contact Ascent Research.
Ascent Research offers comprehensive support for the NR4A1 Knockout Raji Polyclonal Cells, including assay protocols and technical consultation, to facilitate your investigations into NR4A1 biology and therapeutic targeting in B-cell malignancies.
