The CLN5 Knockout Raji Polyclonal Cells represent a heterogeneous population of Raji B lymphocytes engineered using CRISPR/Cas9 to disrupt the CLN5 gene, generating a polyclonal loss-of-function model for lysosomal biology research. This gene-edited polyclonal product preserves the natural genetic diversity of a knockout pool, offering a robust system to study CLN5-associated pathways without the biases of single-cell clones.
The Raji host cell line, derived from a human Burkitt’s lymphoma patient, is an Epstein-Barr virus-positive B lymphocyte model that grows in suspension and expresses characteristic B cell surface markers CD19 and CD20. As a transformed lymphoblast, Raji cells are widely employed to study B cell biology, oncogenic signaling, and hematological malignancies, providing a versatile and well-characterized platform for genetic manipulation.
CLN5 encodes a soluble lysosomal protein that participates in lysosomal degradation and glycosphingolipid metabolism. Within the endolysosomal system, CLN5 forms functional complexes with other neuronal ceroid lipofuscinosis-associated proteins, including PPT1/CLN1, TPP1/CLN2, CLN3, CLN6, and CLN8, and it interacts with LAMP1 and RAB7A to regulate lysosomal trafficking and acidification. Its expression is transcriptionally controlled by the master lysosomal regulators TFEB and MITF in response to lysosomal stress signals, and its disruption impairs downstream cathepsin enzyme activities, blocks autophagic flux as shown by LC3-II and p62/SQSTM1 accumulation, and leads to sphingolipid accumulation, ultimately resulting in the deposition of autofluorescent storage material characteristic of CLN5 disease.
The introduction of CLN5 disruption into the Raji B lymphocyte background creates a human model to dissect the intersection of lysosomal dysfunction and B cell biology. Although CLN5 mutations primarily cause neurodegeneration, this lymphoblastoid system permits investigation of cell-autonomous lysosomal defects in an immune context, potentially uncovering B cell-specific roles for CLN5 in processes such as antigen processing or cytokine secretion. The polyclonal nature avoids clonal artifacts, enabling robust assessment of lysosomal pathway perturbations.
These CLN5 knockout polyclonal Raji cells are ideally suited for interrogating lysosomal biology and neuronal ceroid lipofuscinosis pathomechanisms. They enable Western blotting and RT-qPCR confirmation of CLN5 ablation, immunofluorescence and LysoTracker staining to visualize lysosomal expansion and LC3 puncta, and autophagic flux assays to quantify LC3-II turnover. Cathepsin activity measurements and electron microscopy can further validate storage material accumulation. Consequently, this model supports drug discovery for Batten disease, gene therapy validation, and fundamental studies of lysosomal homeostasis. For technical inquiries or ordering information, please contact Ascent Research.
