The DMD Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Raji B-lymphoblastoid cell line, designed to disrupt the dystrophin-encoding DMD gene. This loss-of-function model eliminates dystrophin protein, including the lymphocyte-specific Dp71 isoform, creating a genetically tractable system for functional studies. The polyclonal composition mirrors diverse editing outcomes, offering a population-level knockout tool for robust biochemical and phenotypic assays in an immunologically relevant background.
The Raji cell line, established from a Burkitt??s lymphoma patient, is an Epstein-Barr virus-positive human B-cell lymphoma model. These lymphoblastoid cells retain key B-cell attributes, including surface immunoglobulin expression, antigen-presenting function, and rapid proliferation, making them ideal for non-muscle dystrophin research. In Raji cells, the DMD gene predominantly expresses the Dp71 isoform, which is implicated in cell adhesion, migration, and receptor-proximal signaling events distinct from the full-length muscle dystrophin.
Dystrophin forms a critical scaffold linking the actin cytoskeleton to the extracellular matrix through the dystrophin-glycoprotein complex (DGC), which includes ??-dystroglycan, ??-dystroglycan, sarcoglycans, syntrophins, dystrobrevin, and nNOS. In lymphocytes, Dp71 modulates focal adhesion and downstream MAPK/PI3K-Akt pathways. CRISPR/Cas9-mediated DMD disruption destabilizes this complex, deregulating calpains, osteopontin (SPP1), and cytoskeletal effectors. Transcription factors MEF2 and MYOD regulate DMD expression upstream, while alternative promoter usage produces the Dp71 isoform dominant in immune cells.
In Raji cells, DMD knockout compromises the actin?Cmembrane linkage, causing impaired adhesion, reduced mechanical stability, and aberrant MAPK signaling. This allows detailed study of dystrophin??s role in B-cell homing, activation, and immunoglobulin production. The model is well-suited to investigate how dystrophin deficiency affects immune synapse formation and cell?Cextracellular matrix interactions, contributing to the understanding of extra-muscular dystrophinopathy manifestations.
These polyclonal knockout cells support a wide array of experimental approaches, including Western blotting and immunofluorescence to examine DGC integrity, RT-qPCR and RNA-seq for DMD isoform profiling, and functional assays such as adhesion, migration, and calcium flux measurements. They serve as a platform for high-throughput screening of therapeutic agents capable of restoring dystrophin expression or mitigating downstream signaling defects. For additional product information, please contact Ascent Research.
