The CD2AP Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the CD2AP gene in a human embryonic kidney background. This product provides a heterogeneous pool of edited cells, enabling robust and flexible analysis of CD2AP-dependent signaling without the constraints of clonal selection. The knockout model is suitable for investigating scaffold protein functions in receptor-mediated endocytosis, cytoskeletal organization, and downstream signal transduction.
HEK293T cells are a widely utilized human embryonic kidney epithelial-like cell line derived from the HEK293 parental line, stably expressing the SV40 large T antigen. This feature permits high-level episomal replication of plasmids containing the SV40 origin of replication, making the line particularly valuable for transient and stable protein expression studies. The epithelial origin and robust transfectability of HEK293T cells provide a relevant context for studying membrane-associated adaptor proteins and cytoskeletal dynamics.
CD2AP encodes a scaffold/adaptor protein linking membrane receptors to the actin cytoskeleton, with roles in adhesion, endocytosis, and signaling. It is activated downstream of nephrin, EGF receptor, and TGF-beta receptors, and interacts with cortactin, CAPZ, PI3K, and CIN85. Through these interactions, CD2AP controls PI3K/AKT, NF-kB, and TGF-beta/SMAD signaling cascades, regulating actin polymerization, receptor internalization, and transcription.
In HEK293T cells, CD2AP disruption impairs receptor-cytoskeletal linkage, altering actin dynamics, adhesion, and growth factor receptor trafficking. The epithelial-like background supports study of podocyte-related signaling mechanisms, despite not fully recapitulating differentiation. The knockout enables dissection of contributions to adhesion complex assembly, endocytic sorting, and modulation of PI3K/AKT and TGF-beta/SMAD pathways, informing kidney filtration and neurodegeneration.
Researchers can employ this CD2AP knockout model in a range of experimental applications, including examinations of cell motility and invasion, co-immunoprecipitation of receptor-cytoskeletal complexes, and quantitative analysis of phospho-signaling networks by western blotting. Commonly used assays such as immunofluorescence staining for actin and cortactin localization, as well as functional endocytosis assays, are readily implemented. The system is valuable for probing mechanisms underlying focal segmental glomerulosclerosis and Alzheimer??s disease. For additional information about validation and technical specifications, please contact Ascent Research.
