The MYPN Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated by disrupting the MYPN gene in the Raji B lymphoblast line. This loss-of-function model enables investigation of MYPN-related processes in a transformed B-cell context. The polyclonal format incorporates multiple editing events, minimizing clonal bias while ensuring robust target-gene ablation. Validated for knockout efficiency, these cells support Western blotting, RT-qPCR, and immunofluorescence assays. Derived from an EBV-positive Burkitt’s lymphoma patient, Raji cells provide a well-characterized platform for immunological and lymphomagenesis research, and MYPN knockout extends this utility to non-muscle cytoskeletal studies.
Raji cells are a human B lymphoblastoid line exhibiting hallmarks of transformed B cells, including high proliferation and B-cell marker expression. Widely used in immunology for B-cell receptor signaling and apoptosis studies, their EBV latency III program influences host transcription and may intersect with cytoskeletal networks. This background offers a physiologically relevant setting to assess MYPN’s roles outside striated muscle, particularly in lymphocyte biology.
Myopalladin (MYPN) is a sarcomeric scaffold linking actin, titin, and nebulin at the Z-disc. It also acts as a mechanosensor by binding the co-transcriptional regulator CARP/ANKRD1, transmitting stretch signals to the nucleus under control of MEF2 and SRF. In the mechanotransduction pathway, MYPN interacts with titin-N2A, ??-actinin, filamin C, ANKRD2, calpain 3, and MURF1, regulating actin dynamics and gene expression. Disruption of MYPN impairs this mechanosignaling axis, with potential consequences in non-muscle cells.
Although MYPN is predominantly studied in muscle, it is expressed in lymphocytes, suggesting roles in B-cell cytoskeletal organization and mechanosensing. B cells require actin remodeling for immune synapse formation and migration. By ablating MYPN in Raji cells, this model allows dissection of non-canonical myopalladin functions in adhesion, activation, and mechanotransduction, potentially uncovering mechanisms relevant to lymphoma. It also serves as a control for CRISPR screens in immune cells.
Applications include Western blotting and RT-qPCR to confirm knockout and assess targets; RNA-seq for transcriptomic profiling; immunofluorescence and phalloidin staining to visualize actin defects; and functional assays such as migration, apoptosis, and flow-based phospho-signaling. This model is suited for screening myopathy phenotypes in non-muscle cells, studying cytoskeletal roles in B lymphocytes, and providing a CRISPR knockout control in immunological studies. For custom applications, contact Ascent Research.
