The DFFA Knockout Raji Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout population derived from the human Raji B lymphoblastoid cell line with targeted disruption of the DFFA gene. This loss-of-function model enables systematic investigation of apoptosis regulation, specifically the inhibition of caspase-activated DNase (DFFB/CAD) by DFFA. The polyclonal format captures a heterogeneous mixture of genetic edits, reflecting population-level responses and minimizing clonal selection bias.
Raji cells originate from a Burkitt??s lymphoma patient and are Epstein-Barr virus (EBV)-positive, exhibiting rapid proliferation and serving as a canonical model for B-cell malignancies. Their transformed phenotype and genomic instability make them particularly relevant for studying the interplay between oncogenic signaling and apoptotic pathways, including the DFFA-DFFB axis.
DFFA encodes the inhibitor of caspase-activated DNase (ICAD), which functions as a key negative regulator of apoptosis. Under homeostatic conditions, DFFA forms a heterodimeric complex with its downstream target DFFB (CAD), keeping the nuclease inactive. In response to apoptotic triggers, Caspase-3 (CASP3) and Caspase-7 (CASP7) cleave DFFA, causing its dissociation from DFFB. Active DFFB then translocates to the nucleus to catalyze internucleosomal DNA fragmentation, a biochemical hallmark of apoptosis. DFFA thus serves as a checkpoint that prevents unscheduled DNA digestion, and its caspase-mediated cleavage is a critical commitment step in programmed cell death.
In Burkitt??s lymphoma, evasion of apoptosis contributes to pathogenesis and treatment resistance. This polyclonal DFFA knockout model in Raji cells allows dissection of how loss of CAD inhibitor function impacts DNA fragmentation efficiency in a B-cell malignancy context. The EBV-positive background may further influence apoptotic signaling, making these cells a valuable resource for exploring virus-host interactions in cell death regulation and for evaluating chemotherapeutic sensitivity in lymphomas.
Research applications include quantitative apoptosis assessment via annexin V staining and TUNEL assays, alongside western blot analysis of DFFA cleavage and caspase activation. Co-immunoprecipitation studies can probe DFFA-DFFB complex dynamics, while flow cytometry for DNA fragmentation and caspase activity measurements offer complementary readouts. These polyclonal knockout cells enable drug-induced apoptosis screening, DNA damage response investigations, and studies of lymphoma resistance mechanisms. For further technical information, please contact Ascent Research.
