The AKT3 Knouckout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the AKT3 gene in Homo sapiens. This product provides a mixed population of A-549 cells with disruption of the endogenous AKT3 locus, facilitating loss-of-function studies without clonal isolation. It is designed for researchers investigating AKT3-dependent signaling networks and biological processes.
The parental A-549 cell line is a human lung adenocarcinoma epithelial line derived from a 58-year-old Caucasian male. It serves as a classical model for alveolar type II pneumocytes and retains key features of lung adenocarcinoma, including the ability to grow as adherent monolayers and respond to oncogenic signaling cues. This background is widely employed in oncology, virology, and respiratory biology research.
AKT3 encodes a serine/threonine kinase that serves as a pivotal effector of the PI3K/AKT/mTOR signaling cascade. Activation occurs through PIP3-mediated membrane recruitment and subsequent phosphorylation by PDK1 at Thr308 and mTORC2 at Ser473. Active AKT3 phosphorylates a range of downstream targets: it inhibits GSK3??, excludes FOXO transcription factors from the nucleus, suppresses TSC2 to activate mTORC1, inactivates PRAS40, and phosphorylates BAD to prevent apoptosis. These events integrate signals from upstream regulators including insulin and IGF, while PTEN opposes pathway flux by dephosphorylating PIP3.
In the A-549 lung adenocarcinoma model, AKT3 disruption enables dissection of isoform-specific contributions to oncogenic signaling. The PI3K/AKT pathway is frequently hyperactivated in non-small cell lung cancer, driving tumor growth, metastasis, and resistance to chemotherapy and targeted agents. By eliminating AKT3 in this polyclonal population, investigators can assess phenotypic changes such as altered proliferation rates, apoptotic thresholds, or migratory capacity, and link them directly to AKT3 function within a relevant tissue context.
These polyclonal knockout cells support a wide range of experimental applications, including signal transduction studies, cancer biology, and drug resistance research. Common assays include western blotting for phospho-AKT (Ser473) to verify pathway blockade, cell viability and proliferation assays, apoptosis detection, migration and invasion assays, and sensitivity profiling with PI3K or AKT inhibitors. Additionally, RT-qPCR can monitor transcriptional changes in AKT target genes, while immunofluorescence can assess altered subcellular localization of FOXO or other effectors. This model is ideal for functional genomics screens and for investigating cross-talk with other signaling pathways. For further information or technical assistance, please contact Ascent Research.
