The MID1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population produced from the human B lymphocyte-derived Raji cell line, designed to eliminate MID1 gene function. The polyclonal format comprises a diverse pool of cells with targeted gene disruptions, providing a loss-of-function model that avoids clonal selection and preserves population heterogeneity. This approach is well-suited for studies requiring robust and reproducible phenotype assessment at the population level, enabling researchers to bypass the limitations of single-cell clones.
The Raji host cell line originates from an EBV-positive Burkitt lymphoma and exhibits characteristics of B lymphoblastoid cells, including the capacity for antibody production and antigen presentation. As an established model for B cell biology and hematological cancers, Raji cells are extensively used to investigate lymphomagenesis, signal transduction, and immune mechanisms. Their EBV genome imparts a unique pathological background that is highly relevant to B cell lymphoma research.
The MID1 gene encodes a microtubule-associated RING-type E3 ubiquitin ligase that, together with its binding partner alpha4 (IGBP1), selectively targets the catalytic subunit of protein phosphatase 2A (PP2A) for ubiquitin-dependent proteasomal degradation. By reducing PP2A levels, MID1 relieves the phosphatase-mediated inhibition of mechanistic target of rapamycin complex 1 (mTORC1), resulting in the hyperphosphorylation of key mTORC1 substrates, including p70S6 kinase (S6K) and eIF4E-binding protein 1 (4E-BP1). This signaling cascade enhances cap-dependent translation initiation through the eIF4E complex. MID1 activity is governed by upstream inputs from Hedgehog/GLI signaling and microtubule dynamics, as well as the PI3K/AKT pathway, and it forms functional interactions with microtubules and the oncogenic transcription factor c-Myc. Thus, MID1 functions as a critical node connecting ubiquitin-proteasome activity with translational control and cytoskeletal organization.
In B lymphocytes, aberrant MID1 expression is implicated in the pathogenesis of B cell lymphomas, where sustained mTORC1 activation promotes uncontrolled proliferation and survival. Raji Burkitt lymphoma cells provide a highly appropriate model to dissect MID1-driven PP2A inactivation and its contribution to lymphomagenesis within an EBV-positive context. This knockout tool enables precise investigation of how MID1 deletion impacts cell growth, apoptosis, antigen presentation, and sensitivity to therapeutic agents such as mTOR inhibitors, offering direct insights into MID1-dependent oncogenic mechanisms.
This polyclonal knockout product is tailored for a diverse array of biomedical applications. It facilitates functional characterization of the MID1/PP2A/mTORC1 pathway using Western blot analysis of phosphorylated S6K and 4E-BP1, PP2A phosphatase activity measurements, co-immunoprecipitation of MID1/PP2A complexes, and quantitative PCR for MID1 transcript levels. The model is also ideal for drug target validation, high-throughput screening of MID1 inhibitors, and exploration of Hedgehog-mediated MID1 regulation in immune cells. Additional experimental approaches include EdU incorporation assays for cell proliferation, Annexin V flow cytometry for apoptosis, ubiquitination assays, and transcriptomic profiling by RNA-sequencing. It further supports studies in hepatocellular carcinoma and pediatric midline malformation disorders. For additional information or tailored support, please contact Ascent Research.
