LAMP2 Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population of the human Raji B lymphocyte cell line, designed for targeted disruption of the LAMP2 gene. LAMP2 encodes a lysosome-associated membrane protein essential for maintaining lysosomal structural integrity and facilitating critical cellular processes, including autophagy and antigen presentation. This polyclonal pool comprises a heterogeneous mix of edited cells, offering a versatile loss-of-function model without clonal selection, and is suitable for investigating LAMP2-dependent mechanisms in a lymphoid context.
The Raji host cell line is derived from a patient with Burkitt lymphoma and is Epstein-Barr virus (EBV)-positive, widely utilized in immunology, virology, and cancer research. As a B lymphocyte model, Raji cells exhibit characteristic features of immune surveillance and antibody production, and they possess robust endolysosomal and autophagic pathways, making them particularly relevant for studying lysosomal biology and MHC class II-mediated antigen processing. Their continuous proliferation and well-characterized signaling networks facilitate reproducible functional assays.
At the molecular level, LAMP2 functions as a key lysosomal membrane protein that stabilizes the lysosomal membrane and coordinates two major pathways: chaperone-mediated autophagy (CMA) and MHC class II antigen presentation. In CMA, the LAMP2A splice variant interacts with the cytosolic chaperone HSPA8 (HSC70) to mediate the translocation of substrate proteins into the lysosomal lumen for degradation. Additionally, LAMP2 contributes to antigen presentation by stabilizing peptide-loading complexes involving MHC class II molecules and the aspartyl protease Cathepsin D within lysosomal compartments. Transcription of LAMP2 is regulated by the basic helix-loop-helix-leucine zipper transcription factors TFEB, MITF, and TFE3, which orchestrate lysosomal biogenesis and autophagy. Downstream, LAMP2 activity influences the function of lysosomal hydrolases and the clearance of autophagic cargo, intersecting with macroautophagy components such as ATG5, ATG7, LC3, and p62/SQSTM1.
In Raji B lymphocytes, disruption of LAMP2 produces a model that recapitulates aspects of lysosomal dysfunction and impaired autophagy, directly relevant to human diseases such as Danon disease, a rare X-linked disorder characterized by cardiomyopathy, myopathy, and intellectual disability. The B cell context enables examination of LAMP2??s dual roles in maintaining lysosomal homeostasis and supporting adaptive immunity, as these cells constitutively express MHC class II molecules and process exogenous antigens. The polyclonal nature of the knockout population provides a spectrum of editing efficiencies, allowing bulk analyses of cellular responses without clonal artifacts, and is particularly advantageous for studying processes that may be masked by clonal heterogeneity.
This knockout product supports a broad range of research applications, including the measurement of autophagic flux using bafilomycin A1 inhibition and LC3 turnover analysis, chaperone-mediated autophagy activity assays, and lysosomal stability evaluations. Researchers can employ Western blotting for LAMP2, LC3, and p62 to assess protein levels, immunofluorescence for lysosomal markers to visualize compartmental integrity, and flow cytometry to quantify MHC class II surface expression. These tools facilitate investigations into the molecular pathology of Danon disease, the regulation of autophagy by TFEB/MITF/TFE3, and the intersection of lysosomal degradation with antigen presentation in B lymphocytes. For additional information or technical support, please contact Ascent Research.
