The AGO3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the HEK293T human embryonic kidney cell line, engineered for targeted disruption of the AGO3 gene. This loss-of-function model provides a versatile tool for investigating Argonaute 3-mediated RNA silencing mechanisms without the constraints of clonal selection.
HEK293T cells are a widely utilized adherent epithelial cell line originating from human embryonic kidney 293 cells, stably expressing the SV40 large T antigen. This immortalized line offers high transfection efficiency and robust protein expression capacity, making it an ideal host for examining molecular interactions and gene regulatory networks. The AGO3 knockout in this background enables direct assessment of AGO3 function in a well-characterized cellular environment amenable to a variety of biochemical and genetic assays.
AGO3 encodes an Argonaute family protein that serves as a core component of the RNA-induced silencing complex (RISC). Following miRNA biogenesis mediated by Dicer, Exportin-5, and TRBP (TARBP2), mature miRNAs are loaded onto AGO3, which subsequently interacts with GW182 (TNRC6A/B/C) to recruit the CCR4-NOT deadenylase complex. AGO3 predominantly mediates miRNA-guided translational repression and mRNA deadenylation-dependent decay, with negligible endonucleolytic cleavage activity compared to its paralog AGO2. Additional interacting factors include PABP and MOV10, which modulate RISC assembly and target recognition, placing AGO3 at the intersection of miRNA processing and post-transcriptional gene silencing.
Disruption of AGO3 in HEK293T cells provides a physiologically relevant context to delineate the functional specialization among Argonaute proteins. Because HEK293T cells endogenously express the full miRNA biogenesis machinery, the polyclonal AGO3 knockout population enables side-by-side comparison with wild-type cells to attribute specific regulatory effects to AGO3. This model is particularly valuable for examining how AGO3 contributes to miRNA destination and target selection, as well as its role in pathways frequently dysregulated in cancer and developmental disorders, where miRNA-mediated control is critical.
Researchers can employ these cells in a wide array of functional assays, including RNA immunoprecipitation to assess RISC composition, luciferase reporter assays to quantify miRNA-mediated repression, and high-throughput approaches such as RNA-seq and miRNA profiling to uncover global changes in gene expression networks. The AGO3 knockout model is well-suited for dissecting miRNA pathway mechanisms, evaluating Argonaute functional redundancy, and exploring the role of miRNA silencing in cancer progression, developmental regulation, and viral infection. For inquiries regarding this product, please contact Ascent Research.
