CASP4 Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal cell population with targeted CASP4 gene disruption in the Raji B lymphocyte suspension line. The polyclonal pool contains a heterogeneous mixture of loss-of-function alleles, generated without clonal isolation, thereby minimizing selection artifacts. The absence of CASP4 protein is expected in the majority of cells, enabling robust investigation of inflammasome and pyroptosis pathways. This product is suited for applications in innate immunity, infectious disease, and oncology research.
Raji is an EBV-positive Burkitt lymphoma cell line that grows in suspension and retains B cell features such as antibody production and antigen presentation. Widely adopted in immunological studies, Raji cells serve as a model for B lymphocyte biology, EBV-driven lymphomagenesis, and host?Cpathogen interactions. Their well-characterized signaling networks and rapid expansion in culture simplify the generation and use of CRISPR knockout derivatives for functional genomics.
Caspase-4 functions as a cytosolic receptor for lipopolysaccharide (LPS), triggering the non-canonical inflammasome pathway. LPS binding induces caspase-4 oligomerization and autocleavage, leading to direct proteolytic activation of gasdermin D (GSDMD). GSDMD N-terminal fragments oligomerize to form membrane pores, executing pyroptosis. In parallel, caspase-4 facilitates NLRP3 inflammasome assembly and caspase-1 activation, which promotes maturation and release of IL-1?? and IL-18. Upstream regulation involves TLR4 and NF-??B, which prime inflammatory gene expression, and interferon-gamma (IFN??) enhances sensitivity. Caspase-4 interacts with NLRP3 and ASC, integrating into a signaling network that includes TRIF, GSDMD, and downstream cytokines.
In Raji cells, CASP4 knockout enables dissection of pyroptosis and inflammasome signaling specifically in B lymphocytes. This model is pertinent to understanding how non-canonical inflammasome activity modulates antibody production, antigen presentation, and EBV-associated oncogenic processes. It also provides a platform for exploring the role of LPS-sensing in B cell responses during sepsis and inflammatory disorders.
Key applications include inflammasome assembly assays (Western blot for cleaved caspase-4 and GSDMD, immunofluorescence for ASC specks), pyroptosis measurement (LDH release, propidium iodide uptake via flow cytometry), and cytokine profiling (ELISA for IL-1?? and IL-18). RT-qPCR can assess transcriptional changes in inflammatory genes. The model is amenable to drug screening for modulators of the non-canonical inflammasome. For inquiries or support, contact Ascent Research.
