AJUBA Knockout HEK293T Polyclonal Cells are a pooled population of human embryonic kidney (HEK) 293T cells subjected to CRISPR/Cas9-mediated gene disruption of AJUBA. This polyclonal knockout model maintains genetic diversity inherent to heterogeneous editing, ensuring robust representation of loss-of-function events. The cells are derived from the widely used HEK293T line and provide an epithelial platform for dissecting AJUBA??s tumor-suppressive and signaling functions.
The host cell line, HEK293T, is a derivative of HEK293 cells stably expressing the SV40 large T antigen. This enables episomal replication of plasmids carrying the SV40 origin of replication, rendering the line exceptionally permissive for transient protein expression and lentiviral production. Derived from human embryonic kidney epithelium, HEK293T cells retain many epithelial characteristics and are routinely employed in studies on signal transduction, protein?Cprotein interactions, and virus?Chost biology.
AJUBA encodes a scaffold that couples cadherin-mediated adhesion to the Hippo tumor suppressor pathway. At adherens junctions, it binds alpha-catenin, beta-catenin, and E-cadherin, and recruits LATS1/2 kinases along with SAV1. This triggers phosphorylation of the transcriptional coactivators YAP1 and TAZ, promoting their cytoplasmic retention and preventing TEAD-driven proliferation. AJUBA also associates with Aurora A kinase to regulate mitotic spindle organization. Upstream, cell?Ccell contact, mechanical stress, and Merlin (NF2) signal through AJUBA. Consequently, AJUBA integrates adhesive and mechanical cues to restrain growth via the Hippo and Wnt pathways.
In HEK293T cells, AJUBA knockout lifts a brake on YAP/TAZ activity even under confluent conditions. This facilitates dissection of AJUBA-dependent Hippo signaling in proliferation and epithelial homeostasis, free from primary cell senescence. The model is apt for studying contact inhibition loss in transformation, as HEK293T already harbors SV40 large T antigen?Cinactivated p53 and Rb. The polyclonal pool thus provides a robust system to examine AJUBA??s role in anchorage-independent growth, migration, and therapeutic response.
Typical applications include Western blotting for phospho-YAP (S127) and immunofluorescence to monitor YAP/TAZ localization, which report Hippo pathway activity. TEAD luciferase assays quantify transcriptional output. Transwell migration and invasion assays assess AJUBA??s effect on cell motility, and co-immunoprecipitation of LATS1 validates complex formation. Flow cytometry for cell cycle analysis and proliferation assays reveal growth control phenotypes. The polyclonal pool is also suitable for lentiviral screens of Hippo modulators. For further information, please contact Ascent Research.
