The AMOT Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of human embryonic kidney cells with targeted disruption of the AMOT gene, which encodes angiomotin. This polyclonal knockout model enables loss-of-function studies of AMOT in a well-characterized epithelial-like cell background.
The host cell line, HEK293T, is a human embryonic kidney epithelial-derived cell line that constitutively expresses the SV40 large T antigen, enabling high transfection efficiency and episomal replication of plasmids containing the SV40 origin of replication. This feature makes HEK293T cells an ideal platform for transient and stable expression studies, as well as lentiviral production, providing a robust context for investigating the functional impact of AMOT ablation.
Angiomotin (AMOT) is a scaffold protein that integrates signals from the Hippo pathway and angiogenic cues to regulate cell migration, polarity, and junction assembly. AMOT acts downstream of LATS1/2 kinases, which phosphorylate AMOT, modulating its interaction with transcription coactivators YAP and TAZ. In the unphosphorylated state, AMOT binds and sequesters YAP/TAZ at tight junctions, preventing their nuclear translocation and transcription of target genes such as CTGF and CYR61. AMOT also interacts with actin filaments and polarity complexes containing Pals1 and Par3 to govern cytoskeletal dynamics and junctional integrity. Upstream regulators include angiostatin and VEGF signaling, while AMOT activity feeds back into actin remodeling and Hippo pathway regulation, positioning it as a critical node in crosstalk between mechanical, angiogenic, and growth-inhibitory signals.
In HEK293T cells, disruption of AMOT is expected to abolish its scaffolding function, leading to unchecked YAP/TAZ nuclear accumulation and constitutive activation of TEAD-mediated transcription. This results in upregulation of pro-proliferative and pro-migratory gene programs, including CTGF and CYR61, and consequent alterations in cell growth and motility. Additionally, loss of AMOT may compromise tight junction integrity and apicobasal polarity, as the interaction network with Pals1 and Par3 is disrupted. This model thus recapitulates key aspects of oncogenic Hippo pathway dysregulation and provides a tractable system to dissect AMOT-dependent control of contact inhibition and epithelial homeostasis.
The AMOT Knockout HEK293T Polyclonal Cells are a versatile tool for investigations into Hippo pathway regulation, angiogenesis, and cell migration. Researchers can employ this model for drug screening of YAP/TAZ or TEAD inhibitors, using TEAD luciferase reporter assays to quantify pathway activity. The polyclonal knockout population is suitable for western blotting to assess YAP/TAZ phosphorylation and total levels, immunofluorescence to monitor YAP subcellular localization, and RT-qPCR for CTGF and CYR61 expression. Transwell migration assays can be used to evaluate the impact on cell motility, while co-immunoprecipitation studies enable mapping of AMOT interactomes. For further technical details, please contact Ascent Research.
