The AKT3 Knockout Ca Ski Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the AKT3 gene in the Ca Ski human cervical carcinoma line. This loss-of-function model enables detailed investigation of AKT3-dependent signaling and cellular processes without pharmacological interference.
Originating from a cervical epidermoid carcinoma, the Ca Ski cell line contains integrated HPV-16 genome, rendering it a paradigm for studying HPV-mediated oncogenesis and therapeutic responses. These cells exhibit dysregulated proliferation and apoptosis resistance, providing a clinically pertinent background for cervical cancer research.
AKT3 is a serine/threonine kinase that functions downstream of PI3K. Activation involves PIP3-mediated membrane recruitment and phosphorylation by PDK1 and mTORC2. Regulatory interactions with PTEN, PHLPP, and HSP90 modulate its activity. Upon activation, AKT3 phosphorylates key effectors: it inactivates GSK3?? and FoxO transcription factors, promotes mTORC1 signaling via TSC2 and PRAS40, and phosphorylates Bad and S6K. These events drive cell survival, proliferation, metabolism, and angiogenesis. Knockout of AKT3 disrupts the PI3K/AKT/mTOR axis, reducing these processes and potentially sensitizing cells to apoptosis.
In the HPV-16-positive Ca Ski background, AKT3 knockout is particularly valuable for dissecting isoform-specific contributions to cervical carcinogenesis. It allows exploration of how HPV oncoproteins may converge with AKT3 signaling, and it facilitates the study of AKT3-dependent chemoresistance mechanisms. This model enables the identification of AKT3-specific substrates and binding partners that may drive malignant phenotypes.
The polyclonal knockout cells support a variety of applications: transcriptomic profiling via RNA-seq or RT-qPCR; biochemical assays such as Western blotting for AKT3, phospho-AKT, and downstream targets like phospho-GSK3?? and phospho-S6; and cellular assays including MTT/CCK-8 proliferation, Annexin V apoptosis, migration/invasion, and colony formation. They are also suitable for drug sensitivity testing with PI3K/AKT/mTOR inhibitors and phospho-signaling antibody arrays. Additionally, these cells can be used to investigate the crosstalk between AKT3 signaling and other pathways affected by HPV oncoproteins. This model is ideal for functional genomics, target validation, and studying HPV-mediated oncogenesis. For further information, please contact Ascent Research.
