The AMOT Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population harboring targeted disruptions of the AMOT gene. This format yields a heterogeneous mix of loss-of-function alleles, providing a representative model for functional studies without the need for clonal expansion. Engineered from the near-haploid HAP1 cell line, these cells facilitate direct assessment of gene function in a simplified genomic background. They are designed for researchers investigating AMOT-dependent processes in cell polarity, junctional integrity, and Hippo pathway regulation.
The HAP1 cell line is a near-haploid human model derived from KBM-7 chronic myeloid leukemia cells. Its primarily haploid karyotype minimizes genetic redundancy, enabling efficient CRISPR-based gene disruption and clear phenotypic outcomes. HAP1 cells grow as an adherent monolayer, making them suitable for imaging and biochemical assays. Their genomic simplicity has established them as a preferred platform for loss-of-function screens and signaling studies.
AMOT (angiomotin) is a tight junction-localized scaffold that regulates Hippo signaling and contact inhibition. It binds YAP and TAZ, preventing their nuclear translocation and TEAD-mediated transcription through interactions with NF2, Pals1, Patj, ZO-1, and F-actin. Angiostatin and TEADs modulate this scaffolding. Under basal conditions, AMOT reinforces the MST1/2?CLATS1/2 kinase cascade, keeping YAP/TAZ phosphorylated and cytoplasmic. Knockout of AMOT allows YAP/TAZ nuclear entry, TEAD activation, and transcription of proliferative and migratory genes. Additionally, AMOT links the actin cytoskeleton to junctional complexes, maintaining epithelial integrity.
In the HAP1 near-haploid environment, AMOT knockout produces unambiguous phenotypes due to the absence of a compensatory allele. This model directly reveals enhanced YAP/TAZ activity, increased TEAD reporter signals, and altered cell migration. It is highly relevant for cancer research??where AMOT loss correlates with tumor progression??and for angiogenesis and vascular malformation studies. Isogenic comparisons with wild-type HAP1 cells enable precise dissection of AMOT-specific functions.
Applications include Western blotting for YAP/TAZ phosphorylation, immunofluorescence to probe junctional architecture, and migration assays. TEAD luciferase assays quantify pathway output, while co-immunoprecipitation maps interaction networks. These polyclonal knockout cells are also ideal for chemical screens targeting Hippo or tight junction modulators. For further information, please contact Ascent Research.
