The CHAC2 Knockout Raji Polyclonal Cells are a population of Raji B lymphoblastoid cells engineered by CRISPR/Cas9-mediated disruption of the CHAC2 gene, yielding a heterogeneous pool of loss-of-function mutants. This polyclonal format avoids clonal biases and provides a robust model to study CHAC2 function in a human B-lymphocyte background. CHAC2 encodes a glutathione-specific gamma-glutamylcyclotransferase that integrates redox status, the unfolded protein response, and Notch signaling, making these cells valuable for dissecting mechanisms in apoptosis and oxidative stress.
The Raji cell line originates from a Burkitt lymphoma patient and is characterized by lymphoblastoid morphology and Epstein-Barr virus positivity. It is an extensively utilized model for B-cell malignancies, humoral immunity, and lymphomagenesis, with well-defined growth in suspension culture and established signaling networks. This background is particularly suitable for gene-editing studies aimed at understanding B-cell transformation, therapeutic resistance, and the contributions of viral oncogenesis to lymphoma development.
CHAC2 catalyzes the cleavage of glutathione into 5-oxoproline and cysteinylglycine, depleting cellular glutathione and fostering oxidative stress. Its expression is induced by ER stress via the ATF4-CHOP axis, linking it to the unfolded protein response. The resulting glutathione depletion triggers caspase-3-mediated apoptosis under chronic stress. CHAC2 also negatively regulates Notch signaling, likely through interaction with the Notch intracellular domain, leading to reduced transcription of targets such as HES1 and HEY1. Thus, CHAC2 integrates ER stress with apoptotic and developmental pathways, with glutathione as both substrate and key redox molecule.
In Raji Burkitt lymphoma cells, CHAC2 disruption enables dissection of crosstalk between oxidative stress, Notch signaling, and survival pathways. As CHAC2 promotes apoptosis under ER stress, knockout cells may show altered sensitivity to stress-inducing agents, providing a model for drug resistance studies. Comparing wild-type and knockout populations helps clarify CHAC2??s role in balancing glutathione metabolism and Notch activity, with implications for B-cell malignancies where these pathways are frequently dysregulated.
These polyclonal cells support diverse assays: glutathione quantification, Western blotting for CHAC2, Notch1, and cleaved caspase-3, RT-qPCR for Notch targets (HES1, HEY1), flow cytometry for Annexin V/PI apoptosis, viability assays with tunicamycin or thapsigargin, and Notch reporter assays. Such experiments facilitate functional characterization of CHAC2 in a lymphoblastoid context. For inquiries or to acquire the CHAC2 Knockout Raji Polyclonal Cells, please contact Ascent Research.
