The ACAP1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human embryonic kidney HEK293T cell line. This product provides a loss-of-function model for the ACAP1 gene, enabling precise investigation of Arf6 GTPase-activating protein function in membrane trafficking and endosomal recycling. The polyclonal format reflects a mixed population of edited cells, offering robust genetic disruption without clonal selection, suitable for population-level functional assays.
HEK293T cells are human embryonic kidney epithelial cells immortalized by stable integration of the SV40 large T antigen. These cells retain functional characteristics of renal epithelial transport and barrier function, and are widely employed as a tractable model system for studying endocytic trafficking, signal transduction, and cell migration. Their epithelial origin and robust growth properties make them an ideal background for analyzing ACAP1-dependent regulation of integrin recycling and cellular dynamics.
ACAP1 encodes an Arf6 GTPase-activating protein that catalyzes GTP hydrolysis on Arf6, a small GTPase pivotal for endosomal membrane traffic. ACAP1 is activated downstream of the EGF receptor and regulated by Rab35 and phosphatidylinositol-4,5-bisphosphate. Upon activation, ACAP1 promotes the conversion of active Arf6-GTP to inactive Arf6-GDP, facilitating the recycling of internalized cargo, including ??1 integrins and EGFR, from peripheral endosomes back to the plasma membrane. ACAP1 physically interacts with clathrin heavy chain and integrin ??1, and functions in concert with Rab35, EHD1, and Rab11 as part of the endocytic recycling machinery. Through this mechanism, ACAP1 modulates actin cytoskeleton remodeling, cell adhesion, and directed cell migration.
In the HEK293T background, disruption of ACAP1 provides a clean system to dissect its role in integrin and receptor tyrosine kinase recycling. Given HEK293T cells?? epithelial phenotype and well-characterized endocytic pathways, the ACAP1 knockout polyclonal population facilitates the study of trafficking-dependent cell adhesion and migration. This model is particularly relevant for cancer metastasis research, where ACAP1-mediated regulation of integrin surface levels and focal adhesion dynamics is critical for invasive behavior.
Researchers can employ this knockout model in a variety of functional assays including integrin recycling assays, transwell migration and invasion assays, and immunofluorescence-based trafficking analysis. Western blotting and RT-qPCR enable validation of ACAP1 loss and monitoring of downstream effectors such as ??1 integrin and Arf6-GTP levels. Co-immunoprecipitation studies can probe ACAP1 interactions with clathrin or Rab35. Additionally, this model enables investigation of EGF receptor signaling dynamics in the absence of ACAP1, offering insights into growth factor?Cdriven cell motility. For additional information, please contact Ascent Research.
