The MTMR12 Knockout Raji Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of Raji B lymphocytes harboring a targeted disruption of the MTMR12 gene. This loss-of-function model provides a genetically defined system for investigating the roles of the MTMR12 pseudophosphatase adaptor in endosomal phosphoinositide metabolism and autophagy regulation. The polyclonal knockout format provides a diverse allelic pool for robust functional analyses without clonal selection bias. These suspension cells retain parental Raji characteristics with abolished MTMR12 expression.
Raji cells are an Epstein-Barr virus (EBV)-positive human B-lymphocyte line derived from a Burkitt lymphoma patient. They retain germinal center B-cell features and active B-cell receptor signaling. EBV latent gene expression modulates apoptosis and proliferation, establishing Raji as a standard lymphoma and B-cell immunology model. The hematopoietic origin and robust growth kinetics of Raji cells facilitate diverse experimental manipulations, including genetic knockout studies.
MTMR12 is a catalytically inactive pseudophosphatase scaffolding the MTM1 phosphatase complex on endosomal and autophagosomal membranes. It directly interacts with MTM1, Beclin 1, and the lipid kinase VPS34 to spatially regulate phosphatidylinositol 3-phosphate (PI3P) pools. Upstream signals from VPS34, mTORC1, and AMPK modulate its scaffolding function, which directs MTM1-mediated PI3P dephosphorylation and controls downstream WIPI2 recruitment and LC3 lipidation. This coordinates autophagosome maturation and endosomal trafficking in response to nutrient and stress cues.
In the Raji lymphoma background, disruption of MTMR12 perturbs the PI3P-dependent endosomal sorting and autophagy pathways that are often dysregulated in B-cell malignancies. EBV-associated lymphomas exhibit altered autophagy and endocytic signaling, processes in which MTMR12 is a critical node. This knockout model thus enables dissection of how pseudophosphatase adaptor-dependent PI3P metabolism influences lymphoma cell growth, survival, and response to microenvironmental cues. The model is also relevant for studying mechanisms underlying centronuclear myopathy and neurodegenerative disorders where MTMR12 dysfunction has been implicated.
This product enables autophagy flux assays (LC3/p62 Western blotting and bafilomycin A1 treatment), endosomal marker immunofluorescence, and PI3P lipid ELISA. Co-immunoprecipitation of the MTM1?CBeclin 1?CVPS34 complex and flow cytometric apoptosis assays are also applicable. Researchers focusing on phosphoinositide signaling, autophagy regulation in B cells, or endolysosomal pathway analysis will find this model a valuable tool for genetic perturbation studies. For further technical details or ordering information, please contact Ascent Research.
