The DUSP11 Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the DUSP11 locus is disrupted across a heterogeneous pool of cells. This format provides a versatile loss-of-function model for investigating DUSP11 biology without the biases introduced by single-cell cloning, ensuring that functional studies capture a broad allelic spectrum.
HEK293T is a widely used human embryonic kidney cell line transformed with adenovirus 5 DNA, characterized by high transfection efficiency and robust protein production. It is a preferred host for viral packaging and cell biology experiments. The line retains intact innate immune signaling pathways, making it particularly suitable for dissecting mechanisms of RNA sensing and interferon responses in a defined cellular background.
DUSP11 encodes an RNA 5′-phosphatase that catalyzes the stepwise removal of the ?? and ?? phosphates from 5′-triphosphorylated RNA, yielding 5′-monophosphorylated RNA. This modification is critical for preventing recognition by the cytoplasmic innate immune receptors RIG-I and MDA5, which specifically detect 5′-triphosphorylated RNA as a pathogen-associated molecular pattern. Consequently, DUSP11 serves as a negative regulator of the RIG-I/MAVS/TBK1/IRF3 signaling axis, limiting the production of type I interferons such as interferon-beta. DUSP11 additionally interacts with the RNA exosome complex to modulate microRNA processing and contributes to the turnover of RNA species. Its expression is induced by type I interferon signaling through the JAK-STAT pathway, establishing a regulatory feedback loop that dampens innate immune activation.
In the HEK293T background, loss of DUSP11 results in the accumulation of endogenous 5′-triphosphorylated RNA, which constitutively activates RIG-I-dependent signaling and downstream interferon responses. This makes the polyclonal knockout cells a powerful system for exploring how alterations in RNA metabolism impact innate immunity and for studying viral evasion mechanisms. The model is also valuable for tumor immunology research, as DUSP11 deficiency may alter the immunogenicity of cells, and for functional genomics screens using CRISPR-based approaches.
Typical assays performed with these cells include RT-qPCR analysis of interferon-stimulated gene expression, RIG-I activation reporter assays, RNA immunoprecipitation, detection of 5′-triphosphorylated RNAs, microRNA expression profiling, and interferon-beta ELISA. Viral infection studies can reveal the dependency of viral replication on DUSP11-mediated RNA modification. The polyclonal nature of the cell population provides a more representative functional landscape, reducing clonal artifacts. For technical inquiries, please contact Ascent Research.
