The AGO4 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from Jurkat human T lymphocytes, designed for the study of AGO4 loss-of-function. This cell pool contains a heterogeneous mix of targeted gene disruptions at the AGO4 locus, providing a model system that avoids clonal bias. The polyclonal nature is suited for experiments where population-averaged effects are informative, such as miRNA pathway analysis and functional screens.
Jurkat cells are an immortalized T lymphocyte line established from an acute T cell leukemia patient. They recapitulate many features of T cell signaling and activation, responding to stimuli like anti-CD3/CD28 antibodies and phorbol esters. Their relevance to leukemia research and ease of genetic manipulation make them a widely used host for studying gene function in T cell biology and oncogenesis.
AGO4 encodes an Argonaute protein integral to the RNA-induced silencing complex (RISC). It binds mature miRNAs and guides post-transcriptional silencing of target mRNAs via cleavage or translational repression. AGO4 function depends on interactions with TNRC6, DICER, TRBP, and miRNA duplexes. The gene is regulated by upstream factors p53, MYC, and NF-??B, and its activity influences downstream targets such as the tumor suppressors PTEN and CDKN1A. Disrupting AGO4 therefore impairs miRNA-mediated gene control.
Loss of AGO4 in Jurkat cells is expected to disrupt miRNA-guided regulation, potentially altering the expression of genes controlling T cell proliferation, survival, and activation. Because miRNAs are often dysregulated in leukemia, AGO4 knockout provides a means to dissect the contribution of RISC-dependent silencing to T cell leukemogenesis. The polyclonal format enables the observation of consensus phenotypes without clonal selection artifacts.
Applications of these cells include investigating miRNA function in T cells, mapping RISC interactions, and performing functional genomics screens. Researchers can validate AGO4 knockout via Western blotting, quantify miRNA and target gene levels by RT-qPCR, assess RISC activity through luciferase reporter assays, and examine transcriptome-wide changes with RNA-seq. Functional assays such as flow cytometry for T cell activation markers, apoptosis, and proliferation assays further characterize AGO4-dependent phenotypes. For more information, please contact Ascent Research.
