The AGO4 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 colorectal adenocarcinoma cell line. This product features disruption of the Argonaute 4 (AGO4) gene, resulting in a pooled heterogeneous culture lacking functional AGO4 protein. The polyclonal format ensures diverse genetic edits while enabling loss-of-function studies of AGO4-dependent post-transcriptional regulation without clonal bias.
The parental HT29 cell line, derived from a female patient, is a hypertriploid human colorectal adenocarcinoma model with mutant p53 and epithelial characteristics. Under specialized culture conditions, HT29 cells can form polarized monolayers and differentiate, serving as an important platform for intestinal epithelial biology and colorectal cancer research. This well-characterized line supports studies on oncogenic signaling, tumor cell differentiation, and therapeutic interventions.
AGO4 is a pivotal component of the RNA-induced silencing complex (RISC) that binds mature miRNAs and siRNAs to guide post-transcriptional gene silencing by promoting mRNA degradation or translational repression. Small RNA loading into AGO4 is orchestrated by Dicer and its partner TRBP, while downstream effector interactions with TNRC6/GW182 proteins recruit deadenylases to target transcripts. AGO4 expression is regulated by transcription factors such as c-MYC and is linked to upstream pathways involving Drosha/DGCR8 and Exportin-5. Consequently, AGO4 ablation in HT29 cells removes a key node in the RNA interference network.
In the HT29 colorectal cancer environment, AGO4 loss disrupts miRNA-mediated control over genes implicated in cell proliferation, apoptosis, and epithelial-mesenchymal transition. The polyclonal knockout population captures a range of mutations, offering a robust average effect that mitigates clonal variability. This model enables dissection of AGO4??s role in modulating colorectal adenocarcinoma phenotypes and provides a tool to explore functional compensation among Argonaute family members.
These polyclonal cells are ideal for applications such as miRNA target identification, RNAi pathway studies, and cancer biology research. Typical assays include Western blotting to verify AGO4 knockout, RT-qPCR to profile target gene expression, miRNA luciferase reporters to assess silencing function, and RNA immunoprecipitation to analyze RISC interactions. Additionally, cell-based assays for proliferation, apoptosis, and migration can reveal phenotypic consequences of AGO4 loss, and drug sensitivity screens may uncover changes in therapeutic response. For further details or to order this product, please contact Ascent Research.
