The CTSA Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the CTSA gene in the human Raji B lymphocyte line. This gene-edited model introduces loss-of-function modifications through CRISPR/Cas9-mediated gene disruption, creating a heterogeneous pool of edited cells suitable for studying CTSA-dependent processes. Unlike monoclonal lines, this polyclonal format retains genotypic diversity, enabling robust phenotypic analysis while avoiding clonal artifacts. The product is intended for advanced investigations in lysosomal biology, sphingolipid metabolism, and immune cell function, providing a versatile tool for disease modeling and drug discovery.
The Raji cell line, established from a patient with Burkitt’s lymphoma, serves as a well-characterized host for this knockout model. These Epstein-Barr virus (EBV)-positive B lymphocytes exhibit continuous proliferation and express key components of the antigen processing and endolysosomal pathways. Their derivation from a lymphoid malignancy makes them particularly relevant for studying intersections between lysosomal function and immune dysregulation. The Raji background supports experiments in B cell receptor signaling, MHC class II-mediated antigen presentation, and lysosomal enzyme trafficking, offering a physiologically meaningful context for CTSA disruption.
CTSA encodes the lysosomal serine carboxypeptidase cathepsin A, which functions as a protective protein for beta-galactosidase (GLB1) and neuraminidase-1 (NEU1). In its role within the lysosomal multienzyme complex, CTSA stabilizes these glycosidases and is essential for the degradation of glycolipids and glycoproteins. Knockout of CTSA disrupts this complex, leading to reduced catalytic activity of GLB1 and NEU1, thereby impairing lysosomal breakdown of sphingolipids and glycoconjugates. This defect reverberates through sphingolipid metabolism and glycosphingolipid metabolism, with downstream consequences for antigen processing and presentation. Transcription factor TFEB, a master regulator of lysosomal biogenesis, acts upstream of CTSA, along with NF-kB and inflammatory cytokines that modulate its expression. CTSA also interacts with LAMP-1 and cathepsin A-binding proteins, influencing lysosomal integrity and function.
In the Raji B cell context, CTSA knockout models key aspects of lysosomal storage disorders such as galactosialidosis, where CTSA deficiency underlies neurodegeneration and immune dysfunction. The disruption of sphingolipid metabolism in these cells can alter lipid raft composition, potentially affecting B cell receptor signaling and antigen presentation via MHC class II molecules. Additionally, impaired lysosomal degradation may lead to accumulation of undigested substrates, triggering cellular stress responses and apoptosis. This model thus provides a relevant platform to dissect how lysosomal dysfunction drives immune dysregulation and contributes to B cell pathogenesis, including lymphomagenesis and inflammation.
The CTSA Knockout Raji Polyclonal Cells are well-suited for a range of research applications, including galactosialidosis modeling, studies of lysosomal enzyme trafficking, and drug screening for compounds that restore lysosomal function. Representative assays include Western blotting for CTSA and GLB1, enzyme activity measurements for beta-galactosidase and neuraminidase-1, immunofluorescence for lysosomal markers such as LAMP1, flow cytometry to quantify surface MHC class II expression, and mass spectrometry-based glycolipid analysis. Gene expression changes can be assessed by RT-qPCR for lysosomal genes downstream of TFEB. For further information or to inquire about this product, please contact Ascent Research.
