The CLMP Knockout Raji Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population of the Raji B lymphoblastoid cell line, harboring targeted disruption of the CLMP (CXADR-like membrane protein) gene. This engineered cell pool offers a loss-of-function model system for dissecting CLMP??s contributions to homophilic cell adhesion, tight junction assembly, and regulation of paracellular permeability. As a polyclonal population, it encompasses a spectrum of editing events, minimizing clonal bias and enhancing the statistical power of downstream functional comparisons to wild-type Raji controls.
The Raji host cell line is an Epstein-Barr virus (EBV)-positive B lymphocyte model derived from a Burkitt lymphoma patient. These suspension-adapted lymphoblastoid cells exhibit key B-cell functions, including antibody production and antigen presentation, and provide a relevant background for studying adhesion-mediated signaling in hematopoietic cells. Raji cells endogenously express components of epithelial junction complexes under certain conditions, making them suitable for interrogating CLMP??s role in lymphocyte adhesion and barrier-related dynamics.
CLMP localizes to tight junctions and engages in homophilic interactions that scaffold with the zonula occludens protein ZO-1 and the transmembrane protein occludin, linking to the cortical actin cytoskeleton via claudins. This molecular architecture is critical for maintaining epithelial barrier integrity. CLMP expression is regulated by pro-inflammatory cytokines TNF-alpha and TGF-beta, and downstream it modulates the localization and stability of ZO-1, occludin, and claudin-1. In the knockout model, disruption of CLMP is expected to perturb these interactions, leading to compromised junctional complex formation and increased paracellular permeability, paralleling observations in intestinal barrier dysfunction.
In the Raji background, CLMP loss-of-function may reveal novel aspects of B-cell adhesion and activation, given the emerging role of tight junction proteins in lymphocyte homing and immune synapse formation. The polyclonal knockout cells facilitate the study of homophilic adhesion outside traditional epithelial contexts, enabling researchers to explore how CLMP-dependent cell-cell contacts influence B-cell behavior, including aggregation, signaling, and crosstalk with other immune cells. This model also aids in modeling diseases associated with barrier breakdown, such as inflammatory bowel disease and obesity, where CLMP has been implicated.
Researchers can employ these polyclonal knockout cells in a range of assays: Western blotting and RT-qPCR for expression analysis, immunofluorescence and flow cytometry for protein localization and surface expression, co-immunoprecipitation for protein-protein interactions with ZO-1 or occludin, and cell aggregation assays to quantify adhesion. Transcriptomic profiling via RNA-seq further enables discovery of downstream pathways altered by CLMP loss. This product is an indispensable resource for advancing tight junction biology, lymphocyte adhesion mechanisms, and barrier function studies. For inquiries, contact Ascent Research.
