The NUDT9 Knockout Raji Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of Raji B lymphocytes carrying a targeted disruption of the human NUDT9 gene. This loss-of-function model is generated through CRISPR/Cas9-mediated gene ablation, resulting in a mixed polyclonal pool with NUDT9 inactivation across the cell population. The knockout pool provides a genetically defined tool for interrogating ADP-ribose metabolism and calcium signaling pathways in a B cell context, without reliance on clonal selection or genetically homogeneous isolates. Researchers are advised that this polyclonal format preserves diverse genetic editing events, facilitating robust population-level functional assessments while mitigating clonal artifacts.
Raji cells are a well-characterized human lymphoblastoid cell line originating from a Burkitt lymphoma patient and immortalized by Epstein-Barr virus transformation. They express wild-type p53 and serve as a critical model for B cell biology, immune responses, and B cell malignancies. The Raji background endows this knockout product with relevance to studies of antigen receptor signaling, lymphomagenesis, and lymphocyte activation, while the intact p53 pathway permits investigations into genotoxic stress responses. These characteristics make Raji cells a preferred host for dissecting molecular mechanisms underlying B cell function and dysfunction.
NUDT9 encodes an ADP-ribose pyrophosphatase that hydrolyzes free ADP-ribose to AMP and ribose-5-phosphate, thereby limiting intracellular ADP-ribose concentrations and restraining TRPM2 cation channel activity. In this polyclonal knockout model, loss of NUDT9 function elevates ADP-ribose levels, which promotes TRPM2 channel opening and calcium influx. Calcium entry subsequently activates CAMK2 and the NF-??B transcription factor, driving transcription of pro-inflammatory mediators such as IL1B and IL6, while also engaging apoptotic effectors like CASP3 and CYCS. Upstream of NUDT9, PARP1 and PARP2 generate ADP-ribose polymers under oxidative stress, which are processed by PARG into free ADP-ribose; NUDT9 thus functions as a counterbalancing hydrolase within this network. Magnesium ions serve as an essential cofactor for NUDT9 catalytic activity.
The NUDT9 knockout in Raji cells accentuates the ADP-ribose/TRPM2/Ca2+ axis in a lymphoid lineage, enabling dissection of how oxidative stress signals translate into calcium-dependent immune cell activation and cell death. Given the roles of ADP-ribose and TRPM2 in ischemia-reperfusion injury, neuroinflammation, and metabolic disorders, this model extends beyond B cell biology to broader pathophysiological contexts. In cancer research, the interplay between ADP-ribose metabolism and p53-mediated apoptosis is of particular interest, and the Raji background preserves relevant apoptotic circuitry. The model thus supports mechanistic studies of inflammation-associated signaling in malignant and non-malignant B cells.
This product is suitable for a wide range of experimental applications, including the investigation of ADP-ribose signaling dynamics, screening of small-molecule modulators targeting NUDT9 or TRPM2, and analysis of oxidative stress-induced apoptosis in B lymphocytes. Relevant assays encompass Western blotting and RT-qPCR for NUDT9 expression validation, mass spectrometry-based quantification of cellular ADP-ribose, calcium influx measurements using Fluo-4 AM, flow cytometric detection of apoptosis by Annexin V/propidium iodide staining, ROS detection with DCFDA, ELISA for IL-1?? and IL-6 secretion, NF-??B luciferase reporter assays, and cell viability assessments by MTT or CellTiter-Glo. For further technical specifications and support, please contact Ascent Research.
