The DIAPH1 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population with targeted disruption of the DIAPH1 gene. This heterogeneous knockout model is derived from HEK293T cells and provides a versatile tool for loss-of-function studies in actin cytoskeleton biology. The pooled format avoids clonal selection artifacts, enabling robust comparisons with wild-type controls in experiments requiring population-level responses.
HEK293T is a transformed human embryonic kidney epithelial cell line stably expressing the SV40 large T antigen. It is widely employed for high-efficiency transient transfection and viral production. The cells maintain epithelial characteristics, including prominent actin stress fibers and focal adhesions, making them suitable for examining formin-mediated cytoskeletal rearrangements.
DIAPH1 encodes a diaphanous-related formin that nucleates and elongates unbranched actin filaments. Inactive DIAPH1 is autoinhibited; binding of GTP-bound RhoA alleviates this autoinhibition, enabling profilin-dependent actin assembly. DIAPH1 interacts with APC, IQGAP1, and microtubule plus-end tracking proteins EB1 and CLIP-170, coordinating actin and microtubule dynamics. Downstream, DIAPH1-driven actin polymerization promotes MRTF-A/SRF transcriptional activity, inducing genes for cell adhesion and migration. Upstream signals include lysophosphatidic acid (LPA) and integrin-mediated adhesion, which converge on RhoA activation. Additionally, DIAPH1 participates in cytokinesis by regulating the contractile ring, and its knockout impairs cell division fidelity.
In HEK293T cells, DIAPH1 knockout disrupts linear actin filament formation, offering a system to dissect formin-specific contributions versus Arp2/3-mediated branching. The model facilitates analysis of adhesion turnover and migration mechanisms relevant to epithelial cell biology and cancer invasion. Although HEK293T are not physiologically matched to inner ear hair cells, the knockout can aid in studying actin pathology related to DFNA1 hearing loss and immunodeficiency.
Typical applications include immunofluorescence with phalloidin to visualize F-actin, western blotting for DIAPH1 and SRF targets, transwell migration assays, and RhoA activation assays. Co-immunoprecipitation with RhoA or profilin can probe disrupted interactions. The cells support screening of compounds targeting Rho-DIAPH1 signaling for anti-metastatic therapy. For customized services or additional information, please contact Ascent Research.
