The MCOLN1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of Raji cells with disrupted MCOLN1, generating a loss-of-function model for the TRPML1 lysosomal cation channel. This knockout model is supplied as a heterogeneous cell pool, avoiding clonal selection bias and enabling study of gene function in a B lymphocyte context. The targeted disruption impairs endogenous TRPML1 expression, providing a key tool for dissecting lysosomal signaling and homeostasis.
The Raji cell line, an EBV-positive Burkitt??s lymphoma-derived B lymphoblast, is a widely used model for B cell malignancies and immune function. These cells exhibit robust proliferation and apoptotic resistance, making them suitable for investigating lysosomal biology in lymphoma. The Raji background provides a lymphoid environment to explore how lysosomal pathways contribute to malignant B cell survival and immune evasion.
MCOLN1 encodes TRPML1, a late endosomal/lysosomal Ca2+ and Fe2+ channel activated by PI(3,5)P2. Upon activation, TRPML1 releases Ca2+ to stimulate calcineurin-mediated dephosphorylation of TFEB, driving its nuclear translocation. Nuclear TFEB induces lysosomal biogenesis and autophagy genes like LC3 and LAMP1, enhancing cellular clearance. TRPML1 interacts with PIKFYVE, ALG-2, mTORC1, and TPC2, and is regulated by starvation, ROS, and lysosomal pH. This signaling axis links lysosomal calcium to mTORC1-autophagy and lysosomal exocytosis pathways.
In Raji B lymphoma cells, MCOLN1 knockout enables dissection of TRPML1??s role in autophagy-dependent survival, lysosomal exocytosis, and stress responses. Lysosomal function is frequently dysregulated in lymphomas, and this model helps elucidate how TRPML1-mediated calcium flux affects mTORC1 signaling, TFEB activity, and pro-survival mechanisms. It provides a platform to study the crosstalk between lysosomal channels and cancer cell homeostasis.
This polyclonal knockout model is ideal for mucolipidosis type IV research, lysosomal storage disorder modeling, autophagy studies, and B cell lymphoma pathobiology. Applications include drug target validation and mechanistic studies using Western blotting for TRPML1, RT-qPCR, immunofluorescence for LAMP1/LC3, lysosomal Ca2+ imaging, autophagy flux assays, and flow cytometry for apoptosis. For inquiries, contact Ascent Research.
