The AKT3 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the AKT3 gene in the human cervical adenocarcinoma HeLa cell line. This polyclonal pool provides a heterogeneous loss-of-function model that enables investigation of AKT3-dependent signaling without the clonal biases inherent in single-cell-derived lines. By leveraging CRISPR/Cas9-mediated gene disruption, this product facilitates robust interrogation of AKT3 function across a diverse cellular background.
HeLa cells are an immortalized adherent epithelial line derived from a HPV18-positive cervical adenocarcinoma and represent one of the most extensively used human cell lines in biomedical research. Their genome is characterized by high aneuploidy and the inactivation of the tumor suppressors p53 and Rb through viral oncoproteins E6 and E7, respectively. These properties establish HeLa as a widely utilized model for studying oncogenic transformation, signal transduction, and therapeutic resistance in an epithelial context.
AKT3 encodes a serine/threonine kinase that serves as a central effector of the PI3K/AKT/mTOR signaling cascade. Upon growth factor stimulation, receptor tyrosine kinases such as EGFR and IGF-1R activate PI3K, which generates PIP3 at the plasma membrane, recruiting AKT3 via its PH domain. Subsequent phosphorylation by PDK1 and mTORC2 fully activates AKT3, enabling it to phosphorylate downstream substrates including GSK3??, FOXO1/3a, TSC2, PRAS40, and BAD. These phosphorylation events drive cell survival, proliferation, and metabolic reprogramming. The pathway is tightly controlled by negative regulators such as PTEN and the phosphatases PP2A and PHLPP. AKT3 also interacts with HSP90 and forms signaling nodes with the mTORC1 complex through TSC2/RHEB regulation.
In the HeLa background, where PI3K/AKT signaling is often hyperactivated due to HPV-driven oncogene expression and genomic instability, AKT3 disruption allows researchers to dissect the isoform-specific contributions of AKT3 to malignancy-associated phenotypes. Given that HeLa cells harbor p53 and Rb inactivation, this knockout model is particularly valuable for interrogating AKT3-mediated survival and proliferation signals independent of these tumor suppressor pathways, offering insights into cancers such as glioblastoma, breast, and prostate carcinomas where AKT3 is frequently implicated.
This polyclonal knockout pool is suited for a range of functional assays including western blotting to monitor AKT3 expression and phosphorylation of downstream targets, cell proliferation and apoptosis analyses, migration and invasion assays, and insulin-stimulated signaling studies. It also serves as an essential tool for isoform-specific pathway dissection, drug resistance profiling, and transcriptomic analyses via RNA-seq. By providing a heterogeneous knockout population, it enables the study of AKT3-dependent phenotypes in a more physiologically representative context. For additional information and technical support, please contact Ascent Research.
