This MICAL2 Knockout Raji Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of Raji B lymphoblastoid cells carrying targeted disruption of the MICAL2 gene. The polyclonal format provides a heterogeneous loss-of-function model that avoids clonal selection bias, enabling robust studies of MICAL2-dependent actin remodeling and semaphorin signaling in a lymphoma-derived background. This knockout pool is designed for functional genomics, drug screening, and mechanistic studies in B-cell biology.
Raji is an EBV-positive B lymphocyte cell line originally established from a Burkitt lymphoma patient. It is extensively used to dissect B cell receptor signaling, antigen processing and presentation, and apoptosis pathways. Raji cells grow in suspension and exhibit stable expression of surface markers such as CD19 and CD20, with rapid proliferation and compatibility with nucleofection methods that facilitate efficient CRISPR-based genome editing. These attributes make Raji a reliable platform for investigating cytoskeletal regulators in hematological malignancies.
MICAL2 belongs to the MICAL family and is a flavoprotein monooxygenase that stereospecifically oxidizes actin methionine-44, leading to F-actin severing and depolymerization. It is activated downstream of semaphorin-plexin signaling, interacting with upstream regulators including Semaphorin 3A, Neuropilin-1, Plexin A1, and Plexin A2. MICAL2 directly binds the cytoplasmic domains of Plexin A1/A2, CRMP2, and Rab35, forming a complex that relays guidance cues to the actin cytoskeleton. This enzymatic activity modulates downstream effectors such as cofilin, Rac1, and CDC42, thereby controlling adhesion, migration, and cell shape. Through these interactions, MICAL2 plays critical roles in axon guidance, immune cell trafficking, and cancer cell invasion.
In Raji B cells, MICAL2 disruption interferes with the semaphorin?Ccytoskeleton axis, potentially altering actin-dependent processes like immune synapse formation, uropod dynamics, and chemotaxis. Given MICAL2??s association with metastasis in multiple carcinomas, this model provides a tool to dissect its role in hematopoietic malignancies. The Raji background enables testing whether MICAL2 promotes invasive behavior through focal adhesion turnover or matrix metalloproteinase activation, and screening for targeted inhibitors.
This polyclonal knockout population supports diverse assays: immunofluorescence with phalloidin for F-actin visualization, transwell migration, Matrigel invasion, and live-cell imaging of actin dynamics. Co-immunoprecipitation can evaluate MICAL2 complexes with Plexin A1 or Rho GTPases; Western blot and RT-qPCR verify gene disruption. Further readouts include cell adhesion on ECM substrates, flow cytometry for integrins, and pyrene-actin polymerization assays. The cells are suitable for anti-metastatic compound screening and cytoskeletal inhibitor testing. Researchers can also utilize these cells to study semaphorin signaling epistasis by expressing exogenous ligands or GTPases. For additional information, contact Ascent Research.
