The EFHD1 knockout Raji polyclonal cells are a CRISPR/Cas9-edited polyclonal population with targeted disruption of the EFHD1 gene in a human B-lymphocyte background. This loss-of-function model enables investigation of EFHD1-dependent molecular processes without introducing specific clonal artifacts, providing a versatile platform for functional genomics in B-cell research. The polyclonal format preserves population-level heterogeneity while abolishing EFHD1 expression, allowing robust assessment of its roles in signaling and apoptosis.
Derived from the Raji B-lymphoblastoid line, a classical Epstein-Barr virus-transformed cell line originating from a Burkitt lymphoma patient, these cells express characteristic B-cell markers such as CD19 and CD20 but lack surface immunoglobulin. They retain key features of B-cell lineage, including antibody-mediated immune functions and properties relevant to immune surveillance, making them a well-established model for studying B-cell malignancies and receptor-driven signaling cascades.
EFHD1 encodes a calcium-binding protein harboring two EF-hand domains that senses intracellular calcium elevations following B-cell receptor (BCR) engagement. Mechanistically, EFHD1 functions downstream of BCR-proximal kinases LYN and SYK, as well as the phospholipase PLC??2, which generates IP3 to release calcium from intracellular stores. Calcium-bound EFHD1 interacts with F-actin and calmodulin to modulate actin cytoskeleton reorganization, while simultaneously binding to BAX and mitochondrial VDAC to regulate mitochondrial outer membrane permeabilization, cytochrome c release, and caspase-3 activation. Upstream regulators include BCR activation, calcium influx, NF-??B, and TNF-alpha, placing EFHD1 at a convergence point for survival and apoptotic signals.
In the Raji cell context, EFHD1 deletion is predicted to disrupt integration of BCR-mediated calcium signals with actin dynamics and mitochondrial apoptosis, potentially impairing normal cytoskeletal responses and apoptotic control. This deficiency can lead to altered cell survival, diminished apoptotic priming, and aberrant activation, mirroring features of Burkitt lymphoma and other B-cell malignancies. Consequently, this knockout model provides a relevant system to dissect the molecular underpinnings of lymphomagenesis and the calcium-dependent checkpoints that govern B-cell fate.
This product supports a range of research applications, including detailed analysis of BCR signaling and apoptosis using Western blotting for EFHD1, BCL2, and cleaved caspase-3, RT-qPCR for BCL2 family gene expression, and flow cytometry with Annexin V staining for apoptosis or Fluo-4 for real-time calcium flux. Further studies of protein interactions are enabled by co-immunoprecipitation of EFHD1 with BAX or calmodulin, while proliferation assays (CFSE) and transcriptome-wide RNA-seq can reveal broader functional consequences. These applications facilitate therapeutic target evaluation in B-cell malignancies and drug response profiling. For inquiries or ordering, please contact Ascent Research.
