AGO4 Knockout Cell Line (HCT 116)

The choice depends on the experimental question. AGO4 (argonaute 4) is one of four Argonaute family proteins (AGO1-4) that load small RNAs to mediate gene silencing. The Knockout line is appropriate for asking whether AGO4 is required for predicted activities — AGO4 is one of four mammalian Argonaute proteins; AGO2 is the principal 'slicer' (endonuclease activity for siRNA-mediated cleavage), while AGO1, AGO3, and AGO4 lack robust slicer activity but participate in miRNA-mediated translation repression. Overexpression is useful for studying AGO4 in heterologous expression contexts. Important consideration: AGO1, AGO2, AGO3 paralog expression analysis aids interpretation given substantial functional overlap — single AGO4 knockout may show modest phenotypes if other AGOs compensate. For colorectal cancer-context Argonaute research, the EDITGENE AGO4 Knockout in HCT 116 enables study of AGO4-specific functions in MSI-H/dMMR colorectal cancer context. Rescue with wild-type AGO4 is the standard specificity control. The knockout is valuable for studying Argonaute family biology, miRNA-mediated regulation, and emerging AGO4-specific functions in cancer biology.

Primary applications: • miRNA-mediated regulation: miRNA target repression analysis in AGO4-null cells. • Argonaute family dissection: AGO1, AGO2, AGO3 expression analysis to interpret AGO4-specific functions. • small RNA loading: small RNA-AGO4 immunoprecipitation analysis. • MSI-H colorectal context: in HCT 116, AGO4's role in MSI-H/dMMR colorectal cancer biology. EDITGENE recommends this model for researchers investigating Argonaute family biology and AGO4 functional dissection.

Yes. AGO4 rescue experiments require attention to Argonaute architecture: • Construct design: use a codon-modified AGO4 sequence with a small C-terminal tag (FLAG, HA). AGO4 has N, PAZ, MID, and PIWI domains (canonical Argonaute architecture but lacking robust slicer activity) — preserve all elements. • small RNA-binding-deficient rescue: PAZ/MID domain mutations disrupt small RNA loading. • Functional readout: rescue should restore AGO4-dependent miRNA target repression. HCT 116 transduces efficiently with lentivirus and supports stable rescue line generation.