The EFHD2 Knockout HEK293T Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout pool of human HEK293T cells, targeting the EFHD2 gene. This heterogeneous population offers a loss-of-function model for studying EFHD2-dependent processes without clonal bias. Suitable for pooled screens, biochemical assays, and comparisons with wild-type controls, the model enables dissection of EFHD2??s roles in calcium signaling and actin dynamics.
HEK293T cells are human embryonic kidney epithelia immortalized with adenovirus E1A and SV40 large T antigen. These adherent, epithelial-like cells are widely used for recombinant expression, viral packaging, and transfection-based assays due to their high transfectability and robust growth. Despite their non-lymphoid origin, HEK293T cells harbor calcium signaling components and actin regulators, providing a tractable system for ectopic expression of relevant pathway members to reconstitute EFHD2 activities.
EFHD2 (Swiprosin-1) is an EF-hand calcium-binding protein modulating B-cell receptor (BCR) signaling and actin dynamics. It is activated upstream by BCR engagement (CD79A/B), interferon-gamma, and NF-kB pathway activation. EFHD2 promotes apoptosis through interaction with SWAP-70, leading to F-actin polymerization changes, caspase-3 activation, and upregulation of pro-apoptotic Bim. It may suppress NF-kB activity. EFHD2 also directly binds F-actin and filamin A to regulate cytoskeletal organization. Downstream of BCR, calcium mobilization via SYK, BTK, and PLC??2 converges on EFHD2, linking calcium flux to cytoskeletal and apoptotic outcomes.
In HEK293T cells, EFHD2 knockout enables study of calcium-responsive functions outside the lymphocyte context. Although lacking endogenous BCR, HEK293T can be engineered to express BCR components, allowing reconstitution of EFHD2 interactions with SWAP-70 and F-actin. The polyclonal model is ideal for analyzing calcium flux, actin filament architecture, and apoptosis. Given robust NF-kB activity in HEK293T, the knockout provides a clean system to test EFHD2??s modulatory role. Structure-function studies can be performed by reintroducing wild-type or mutant EFHD2 to map calcium-binding and interaction domains.
Applications include western blotting and RT-qPCR for expression analysis, immunofluorescence for actin cytoskeleton visualization, apoptosis assays (Annexin V, caspase-3), NF-kB reporter systems, co-immunoprecipitation for protein complexes, and calcium flux measurements. These applications address research areas such as autoimmune disease, lymphomas, and cancer where EFHD2 dysregulation is implicated. For further details and ordering, contact Ascent Research.
