The AGFG1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the human AGFG1 gene has been disrupted. This loss-of-function model enables targeted investigation of AGFG1??s roles in nucleocytoplasmic transport and endosomal trafficking within a host cell line widely used for protein expression and lentivirus production.
HEK293T is a human embryonic kidney epithelial cell line that stably expresses the SV40 large T antigen, allowing episomal replication of plasmids with SV40 origin. It is characterized by high transfection efficiency, robust recombinant protein expression, and adherent growth. These features make HEK293T a standard platform for genetic manipulation, lentiviral packaging, and functional assays requiring efficient gene delivery.
AGFG1 encodes a nucleocytoplasmic shuttle protein that acts as a cofactor for HIV-1 Rev-dependent nuclear export of unspliced viral RNA via CRM1. Simultaneously, AGFG1 functions as a GTPase-activating protein for ARF6, downregulating ARF6 to regulate endosomal sorting, clathrin-mediated endocytosis, and actin cytoskeleton remodeling. AGFG1 interacts with Rev, CRM1, nucleoporins, ARF6, clathrin, actin, and Rab GTPases such as Rab5 and Rab11. Its activity is modulated by upstream signals from Rev, the epidermal growth factor receptor (EGFR), and integrin pathways, linking viral and cellular processes.
Disruption of AGFG1 in HEK293T cells provides a defined system to study its dual function without background HIV-1 replication. Co-expression of Rev and viral RNA reporters permits analysis of CRM1-mediated export defects. Additionally, HEK293T??s well-characterized endocytic and actin machinery allows detailed examination of ARF6-dependent trafficking and actin dynamics in the absence of AGFG1. Knockout cells are expected to show impaired Rev-dependent RNA export and dysregulated ARF6 activity, resulting in aberrant endosomal cargo sorting and actin polymerization, making them a valuable tool for dissecting these interrelated pathways.
Key applications include investigating host factors in HIV-1 replication, endosomal trafficking mechanisms, ARF6 signaling, actin cytoskeleton regulation, and pooled CRISPR screening. Representative assays such as RNA export assays, endocytosis assays, actin polymerization assays, GTPase activity measurements, western blotting, immunofluorescence, co-immunoprecipitation, and flow cytometry are compatible with this polyclonal knockout population. These cells support functional complementation studies and high-content phenotypic screens. For technical inquiries or ordering information, please contact Ascent Research.
