The DIAPH2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T human embryonic kidney cell line. This product provides a loss-of-function model through targeted disruption of the DIAPH2 gene, enabling researchers to investigate the functional consequences of DIAPH2 depletion in cellular processes.
The host HEK293T cell line is an adherent, immortalized line originating from human embryonic kidney cells that stably expresses the SV40 large T antigen. This background confers high transfection efficiency, robust protein expression, and the capacity to produce recombinant viruses, making HEK293T a versatile platform for molecular and cell biology studies.
DIAPH2 encodes a member of the formin family of actin nucleation factors that translate Rho GTPase signals into polarized actin filament assembly. The protein is activated downstream of the Rho GTPases RhoA, Rac1, and Cdc42, which direct DIAPH2 to promote the polymerization of linear actin filaments, leading to the formation of stress fibers and filament bundles. DIAPH2 achieves this through interactions with profilin-actin complexes and additional partners including APC and CLIP-170. These molecular interactions place DIAPH2 at a critical node within the Rho signaling module, directly linking upstream GTPase activation to downstream actin cytoskeleton reorganization.
In the HEK293T cellular environment, DIAPH2 knockout disrupts these actin-dependent processes, leading to impaired cell migration, adhesion, and cytokinesis. The loss of functional DIAPH2 compromises the ability of cells to reorganize their actin cytoskeleton in response to extrinsic cues such as growth factors or extracellular matrix interactions. Consequently, this polyclonal knockout population provides a physiologically relevant model for investigating the molecular underpinnings of cell motility, morphological transitions, and planar cell polarity.
This model is suitable for a wide range of experimental applications, including the study of cytoskeletal dynamics, Rho GTPase signaling networks, and the mechanistic basis of diseases associated with DIAPH2, such as X-linked intellectual disability and cancers exhibiting aberrant DIAPH2 expression. Compatible assays include immunofluorescence microscopy to visualize actin architecture, western blot analysis to confirm DIAPH2 protein depletion, wound healing and transwell invasion assays to assess migratory capacity, co-immunoprecipitation to map protein interaction networks, Rho GTPase activation assays, and RT-qPCR to profile gene expression changes. For further technical specifications, contact Ascent Research.
