MYO5B Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal pool derived from the human Raji B lymphoblastoid cell line, with disruption of the MYO5B gene. This loss-of-function model enables study of MYO5B-dependent processes in a hematopoietic context. The polyclonal format retains genetic heterogeneity, suitable for initial functional screening. The cells are generated via CRISPR/Cas9-mediated gene disruption, abolishing MYO5B protein expression for investigating intracellular trafficking and cell polarity.
The Raji host cell line, an EBV-positive B lymphoblastoid line from Burkitt lymphoma, models B-cell malignancies, immune function, and antigen presentation, expressing B-cell receptors and MHC class I/II. Its suspension growth and well-characterized signaling facilitate studies of endosomal recycling, immune synapse formation, and migration, making it ideal for dissecting MYO5B in B-cell biology.
MYO5B encodes an actin-based motor transporting Rab11a-positive recycling endosomes to the plasma membrane, critical for polarized delivery of cargo proteins like CFTR, DPP4, and E-cadherin, essential for microvilli and membrane homeostasis. It directly interacts with Rab11a, Rab8a, Rab11-FIP2, actin, calmodulin, and SNARE proteins. Upstream regulators include calcium/calmodulin, Src kinases, and PAK, with downstream effectors in cell polarity and adhesion. The Rab11a?CMYO5B?CRab11-FIP2 axis coordinates with sorting nexins to control surface composition.
In Raji cells, MYO5B knockout disrupts apical recycling, reducing surface expression of BCRs and MHC class I, impairing antigen presentation and possibly immune synapse formation. This loss of polarized trafficking affects actin organization and cell migration, relevant for lymphocyte homing and invasion. The model thus enables study of MYO5B in B-cell malignancies, including drug resistance linked to endosomal trafficking, and for testing therapeutic interventions targeting endosomal recycling in lymphoma.
Researchers can employ these cells in Western blotting, RT-qPCR to confirm gene disruption, flow cytometry to quantify surface receptors (e.g., BCR, MHC class I), transferrin recycling assays, transwell migration, and immunofluorescence for actin and recycling endosomes. Co-immunoprecipitation with Rab11a can probe interaction defects. These applications support studies of B-cell receptor recycling, lymphocyte migration, and microvillus inclusion disease-like phenotypes. For further technical details or to discuss custom configuration options, please contact Ascent Research.
