The AGO2 Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely utilized HEK293T cell line, engineered to disrupt the AGO2 gene. This loss-of-function model enables systematic interrogation of AGO2-dependent gene silencing mechanisms without the selection of a single clonal isolate, thus preserving population-level heterogeneity and minimizing clonal biases. The polyclonal format is particularly suited for pooled functional genomics screens and for experimental designs where representation of diverse editing outcomes is desired, providing a robust tool for dissecting RNA interference pathways.
The host HEK293T cell line is an immortalized human embryonic kidney cell line that stably expresses the SV40 large T-antigen, conferring high plasmid amplification and exceptional transfection efficiency. These cells exhibit an adherent epithelial morphology and are a gold standard for transient and stable protein production, lentiviral packaging, and reporter-based assays. The HEK293T background offers a well-characterized molecular landscape, facilitating straightforward interpretation of phenotypic changes upon AGO2 disruption and enabling efficient re-expression or rescue experiments using transfected constructs.
AGO2 (Argonaute 2) is the central catalytic engine of the RNA-induced silencing complex (RISC), where it functions as the slicer endonuclease and orchestrates miRNA- and siRNA-guided post-transcriptional gene silencing. It is regulated by upstream factors including Dicer, TRBP (TARBP2), and AKT-mediated phosphorylation, and is modulated by signals from EGFR and PTEN. Upon loading of mature small RNAs, AGO2 interacts with GW182 (TNRC6A), HSP90, and poly(A)-binding protein PABPC1 to repress translation and promote deadenylation of target mRNAs. Its downstream targets encompass critical transcripts such as MYC, CCND1, BCL2, PTEN, and RAS, placing AGO2 at a nexus of proliferation, apoptosis, and oncogenic signaling. The canonical pathway progresses from pri-miRNA processing by Drosha?CDGCR8 to pre-miRNA export via Exportin-5, followed by Dicer?CTRBP processing and RISC assembly, highlighting the sequential action of these components.
Disruption of AGO2 in HEK293T cells abrogates RISC-mediated silencing, providing a clean background to assign miRNA function, validate direct miRNA?CmRNA interactions, and explore the contribution of AGO2 to cellular phenotypes. Given the HEK293T line??s tractability, this knockout model is invaluable for studying cancer-relevant processes where AGO2 expression or activity is dysregulated, such as in colorectal and gastric cancers, as well as for investigating neurodevelopmental disorders and host?Cviral interactions that depend on RNAi machinery. The polyclonal nature ensures that no single off-target editing event dominates, lending robustness to pooled loss-of-function analyses.
This AGO2 knockout product is suitable for a broad array of downstream applications, including dual-luciferase reporter assays for miRNA target validation, RT-qPCR and RNA-seq to assess target mRNA regulation, co-immunoprecipitation of RISC components, and Western blotting or immunofluorescence to monitor protein-level changes. It can be employed in genome-wide CRISPR screens to identify synthetic lethal interactions or miRNA-dependent vulnerabilities. The combination of a defined knockout background and HEK293T handling advantages makes these cells a versatile platform for both fundamental RNA biology research and applied drug discovery efforts. For further information or to inquire about this product, please contact Ascent Research.
