The MLKL Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human Raji B lymphocyte cell line, designed to disrupt the MLKL gene. This product offers a heterogeneous pool of MLKL-deficient cells, providing a robust loss-of-function model for necroptosis research without clonal artifacts. The genetic modification was achieved through targeted CRISPR/Cas9-mediated gene editing, resulting in disruption of endogenous MLKL expression.
Raji is an EBV-transformed B lymphoblastoid cell line from Burkitt’s lymphoma, widely used as a B lymphocyte model in immunology and cancer research. These suspension cells retain B-cell surface markers and oncogenic features, making them suitable for studying cell death pathways in hematological malignancies and for dissecting necroptotic signaling within a lymphoma context.
MLKL is the terminal effector of the necroptosis pathway. Upon phosphorylation by RIPK3, activated downstream of death receptors such as TNF-R1 by ligands including TNF, FASL, and TRAIL, MLKL oligomerizes and translocates to the plasma membrane to form lytic pores. This results in membrane rupture, cell lysis, and DAMP release. MLKL interacts with RIPK3, HSP90, PEF1, and ALIX, and its activity is modulated within a multiprotein complex that includes RIPK1, FADD, Caspase-8, and CYLD. The knockout cells allow specific interrogation of MLKL-dependent necroptotic events.
In Raji lymphoma cells, disrupting MLKL provides a controlled system to explore necroptosis mechanisms relevant to cancer biology. This model can be used to assess the contribution of necroptosis to drug-induced cell death, to study the crosstalk between necroptosis and inflammatory pathways, and to investigate how DAMP release from necroptotic cells influences the tumor microenvironment. The B-cell malignancy background is especially informative for evaluating necroptosis as a therapeutic vulnerability or resistance mechanism.
Typical applications include western blot analysis of phospho-MLKL and pathway components, propidium iodide uptake and LDH release assays to measure membrane permeabilization, and co-immunoprecipitation of MLKL-RIPK3 complexes. Immunofluorescence can track MLKL translocation, while the cells serve as controls in necroptosis inhibitor screening. The model also supports DAMP release studies and immunogenic cell death research. For technical inquiries, please contact Ascent Research.
