The AGO2 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HeLa cervical adenocarcinoma cell line, featuring targeted disruption of the human AGO2 gene. This product provides a heterogeneous pool of edited cells with loss-of-function mutations in AGO2, enabling loss-of-function studies without clonal selection. The polyclonal format preserves population-level diversity, making it suitable for molecular and biochemical assays where knockout efficiency is verified at the population level and minimizing clone-specific artifacts.
The host HeLa cell line is a widely used HPV-18-positive cervical adenocarcinoma model, characterized by functional inactivation of the tumor suppressors p53 and Rb via the viral oncoproteins E6 and E7. This well-established immortalized line supports robust HPV-18 oncogene expression and recapitulates key features of cervical carcinogenesis. Its rapid proliferation, ease of culture, and extensive molecular characterization make HeLa cells a standard platform for studying gene function in a cervical cancer context and for dissecting oncogenic signaling networks.
AGO2 encodes Argonaute-2, the core catalytic component of the RNA-induced silencing complex (RISC) that possesses slicer endonuclease activity essential for miRNA-guided mRNA cleavage and translational repression. AGO2 directly binds mature miRNAs and, together with accessory proteins such as DICER1, TNRC6/GW182, HSP90, PACT (PRKRA), and TRBP (TARBP2), mediates post-transcriptional regulation by forming RISC on target mRNAs. Upstream regulatory inputs include transcriptional activation by MYC and E2F1, phosphorylation by Akt1 and MAPKAPK2, and signaling through EGFR and HSP90 chaperone pathways. Primary downstream targets encompass miRNA-regulated transcripts such as PTEN, CDKN1A, MYC, and E2F1, thereby controlling cellular processes like proliferation, apoptosis, and differentiation through sequence-specific mRNA deadenylation, decay, or translational inhibition.
In the HeLa model, AGO2 knockout interrogates the interplay between miRNA-mediated gene silencing and HPV-driven oncogenesis. Since HeLa cells express viral E6/E7, which inactivate p53 and Rb, loss of AGO2 disrupts regulatory networks that may further alter expression of key tumor suppressors and oncogenes, offering insights into cervical cancer and other RISC-associated malignancies such as hepatocellular carcinoma. This model is valuable for dissecting how AGO2-dependent silencing impacts viral pathogenesis and the broader miRNA regulome in a transformed epithelial background.
These polyclonal knockout cells are optimized for a range of functional assays, including dual-luciferase reporter systems to quantify miRNA activity, RNA immunoprecipitation (RIP) to monitor RISC assembly, and global approaches such as RNA-seq or small RNA sequencing to assess transcriptome-wide silencing defects. They support mechanistic studies of miRNA-guided mRNA decay and translational regulation, functional genomics screens, and cancer biology research exploring AGO2-dependent growth or apoptotic phenotypes. For additional technical details, validation data, or ordering information, please contact Ascent Research.
