The AGO1 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population engineered for the targeted disruption of the human AGO1 gene in the HEK293T host background. This polyclonal knockout pool provides a heterogeneous population of cells carrying diverse loss-of-function mutations at the AGO1 locus, enabling robust functional studies without clonal isolation. The product serves as a versatile loss-of-function model for dissecting the roles of Argonaute 1 in post-transcriptional gene regulation.
The HEK293T cell line is a human embryonic kidney epithelial derivative widely utilized in biomedical research for its high transfection efficiency, robust protein expression, and capacity for viral production. Stably expressing the SV40 large T antigen, HEK293T cells support episomal replication of plasmids carrying the SV40 origin, making them a preferred system for transient and stable expression studies. Their epithelial morphology and well-characterized signaling networks render them ideal for investigating RNA interference mechanisms, gene regulation, and disease-associated pathways.
AGO1 is a core catalytic component of the RNA-induced silencing complex (RISC), directly binding mature microRNAs (miRNAs) and mediating sequence-specific mRNA cleavage and translational repression. It functions downstream of miRNA processing by Dicer and TRBP, integrating into RISC along with cofactors such as HSP90, PACT, and MOV10. AGO1 interacts with TNRC6A (GW182) to recruit deadenylase complexes, promoting target mRNA destabilization. The protein is regulated by ATP-dependent conformational changes and chaperone activity, and its precise miRNA:target recognition is essential for post-transcriptional silencing networks involving targets like oncogenes and neurodevelopmental regulators.
In the HEK293T epithelial context, AGO1 disruption abolishes its contribution to miRNA-guided RISC activity, permitting dissection of AGO1-specific versus redundant functions shared with other Argonaute proteins such as AGO2. The polyclonal knockout population is particularly suited for examining how AGO1 loss alters miRNA processing dynamics, target mRNA stability, and translational control in an immortalized kidney-derived line. This model is relevant for studying AGO1-associated pathologies including cancer and AGO1-related intellectual disability syndrome, as HEK293T cells retain key pathways that intersect with miRNA-mediated gene silencing.
Researchers can employ these knockout cells in a breadth of functional assays: RNA immunoprecipitation (RIP) for RISC component profiling, small RNA sequencing to assess miRNA loading imbalances, luciferase reporter assays to quantify target suppression, and co-immunoprecipitation to map residual protein interactions. Applications extend to miRNA pathway analysis, cancer gene regulation studies, and neurodevelopmental disorder modeling, where characterizing deregulated mRNA targets is critical. Western blotting and RT-qPCR enable validation of AGO1 knockout and downstream gene expression changes. For further technical details or to discuss custom applications, please contact Ascent Research.
