The CISD2 Knockout Raji Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population genetically modified to disrupt the CISD2 gene in the human Burkitt lymphoma-derived Raji cell line. This polyclonal pool contains a heterogeneous mixture of knockout alleles generated by non-homologous end joining after Cas9-mediated double-strand breaks, yielding a loss-of-function model suitable for pooled knockout studies. The product is designed for researchers investigating the role of CISD2 in B-lymphocyte biology, autophagy, calcium signaling, and mitochondrial function without the bias of single-cell clonal effects.
Raji cells are an Epstein-Barr virus (EBV)-positive B-lymphocyte cell line originally isolated from a patient with Burkitt lymphoma. They grow in suspension and maintain features of mature B cells, including surface immunoglobulin expression and active signaling through B-cell receptor pathways. EBV latency type III in Raji cells establishes a survival program that frequently modulates apoptosis and autophagy, making this background particularly relevant for evaluating how CISD2 knockout reshapes prosurvival networks. The cells are widely employed as a model for lymphomagenesis, immune cell signaling, and viral latency.
CISD2 (also called NAF-1) is an iron-sulfur cluster protein located on the endoplasmic reticulum (ER) and outer mitochondrial membrane. It functions as a critical regulator of autophagy, calcium homeostasis, and redox balance by interacting with BCL2 at the ER and with the IP3 receptor (ITPR1). Under normal conditions, CISD2 reinforces the BCL2-Beclin1 interaction, thereby restraining autophagy initiation. Loss of CISD2 disrupts this complex, freeing Beclin1 to activate the autophagy machinery, as evidenced by increased LC3 lipidation and autophagic flux. Concurrently, CISD2 knockout alters IP3 receptor-mediated calcium release from the ER, perturbing mitochondrial calcium uptake and elevating reactive oxygen species (ROS). These dual effects sensitize cells to oxidative stress and apoptosis, linking CISD2 to p53-mediated stress responses and BCL2 family-regulated cell death.
In the Raji B-cell context, CISD2 knockout provides a powerful system to dissect autophagy-dependent survival mechanisms in lymphoma. The interplay between EBV latency, autophagy, and apoptosis is central to leukemogenesis, and CISD2 disruption can shift this balance toward cell death. Because Raji cells express high levels of BCL2 and are primed for calcium-mediated signaling, the knockout allows direct assessment of how CISD2 modulates the BCL2-Beclin1 axis and calcium dynamics in a malignancy-relevant setting. This model is particularly suited to exploring whether CISD2 loss sensitizes lymphoma cells to chemotherapeutic agents or BH3 mimetics, as well as studying ER-mitochondria communication under metabolic stress.
Typical applications include autophagy flux monitoring via LC3 turnover and p62 degradation in the presence of bafilomycin A1, measurement of cytosolic and mitochondrial calcium using fluorescent reporters, and quantification of ROS with probes such as H2DCFDA. Co-immunoprecipitation can verify disrupted CISD2-BCL2 and CISD2-ITPR1 interactions. Apoptosis assays (annexin V/PI staining) and mitochondrial membrane potential dyes (JC-1) further define the phenotypic consequences of knockout. This model also enables screening of compounds that may bypass CISD2 deficiency, such as those targeting BCL2 or enhancing ATG7-mediated autophagy. For technical specifications or ordering support, please contact Ascent Research.
