ARF4 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the ARF4 gene. This loss-of-function model enables investigation of ARF4-dependent processes in a human embryonic kidney epithelial background. The polyclonal format preserves genetic heterogeneity and avoids clonal artifacts, offering a robust tool for trafficking studies. The HEK293T host line ensures high transfection efficiency and rapid growth.
HEK293T cells are derived from HEK293 by stable expression of SV40 large T antigen, providing a versatile platform for recombinant protein expression and viral production. Their epithelial origin and active secretory pathway make them ideal for examining COPI-mediated retrograde transport and endosomal recycling. These cells are well-suited for expressing and trafficking exogenous cargoes such as GPCRs and rhodopsin.
ARF4 is a small GTPase that regulates COPI-mediated retrograde transport from the Golgi to the ER and endosomal sorting. It is activated by GEFs including GBF1 and BIG1, and upon GTP binding, recruits the coatomer complex to Golgi membranes to initiate vesicle formation. ARF4 interacts with GGA adaptors, Arfaptin, and the ciliary effector RP2, linking it to ciliogenesis. Downstream targets include coatomer subunits COPA and COPB1, while upstream regulators include PI4KIII?? and ARL3. Key pathway components are GBF1, ARF4-GTP, coatomer, and ArfGAP1.
In HEK293T cells, ARF4 knockout disrupts COPI trafficking, affecting Golgi-to-ER retrieval and endosomal recycling. This provides a physiologically relevant model to study membrane dynamics. Given HEK293T’s widespread use for ectopic expression, the knockout facilitates analysis of ARF4-dependent trafficking of overexpressed proteins. The polyclonal design reflects population-level effects, mitigating clonal variations.
These cells support applications in intracellular trafficking, ciliogenesis, cancer cell migration, and drug discovery. Compatible assays include immunofluorescence, GTPase activation, co-immunoprecipitation, vesicle trafficking, complementation rescue, and Sanger sequencing. They are valuable for loss-of-function screens and validation of ARF4 interactors. For further details, contact Ascent Research.
