The AKT3 Knockout 786-O Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the AKT3 gene in the 786-O human clear cell renal cell carcinoma cell line. This polyclonal pool represents a heterogeneous loss-of-function model, offering a genetically diverse background suitable for evaluating AKT3-dependent phenotypes without clonal selection biases. The knockout enables loss-of-function studies in a cancer-relevant epithelial context.
The 786-O host cell line was established from a primary clear cell adenocarcinoma of the kidney in a human male patient. These cells exhibit epithelial morphology and retain characteristics of renal cell carcinoma, including aberrant activation of hypoxia-inducible factor pathways and angiogenic signaling. As a widely used model for studying kidney cancer biology, 786-O cells facilitate investigation of tumor growth, metastasis, and therapeutic responses in vitro and in vivo.
AKT3 is a serine/threonine kinase that acts as a critical effector of the PI3K pathway. Upon activation by upstream regulators including PI3K, PDK1, and mTORC2 downstream of growth factor receptors, AKT3 phosphorylates multiple substrates to promote cell survival, proliferation, and metabolism. Key downstream targets include TSC2, GSK3??, FOXO transcription factors, BAD, p27Kip1, and MDM2, collectively driving mTORC1 activation, survival, and proliferation. Negative regulation is exerted by the tumor suppressor PTEN and by phosphatases such as PP2A and PHLPP. AKT3 also interacts with chaperone HSP90 and the scaffold THEM4. Its signaling integrates via the PI3K-Akt-mTOR and FoxO pathways, coordinating cellular responses to growth and stress signals.
Disruption of AKT3 in 786-O cells ablates AKT3-specific signaling, which is particularly relevant given PI3K-Akt pathway hyperactivation in clear cell renal cell carcinoma. Loss of AKT3 function impairs mTORC1-dependent growth, sensitizes cells to apoptosis via FOXO-driven transcription, and reduces GSK3?? inhibition, potentially attenuating proliferation. This polyclonal knockout model enables dissection of AKT3-mediated mechanisms in renal cancer, including drug resistance and tumor maintenance. The 786-O background provides a clinically relevant platform to study interplay between AKT3 inactivation and other oncogenic drivers such as VHL loss.
Applications include signal transduction studies, drug resistance profiling, apoptosis and proliferation assays, and renal cell carcinoma modeling. Researchers can employ western blotting to verify AKT3 knockout and assess phosphorylation status of downstream targets like GSK3?? and FOXO. Functional assays such as proliferation assays (e.g., MTT, BrdU), apoptosis detection (Annexin V staining), migration/invasion assays (Boyden chamber), and soft agar colony formation can be utilized to evaluate phenotypic consequences. RT-qPCR can further quantify transcriptional changes in target genes such as p27 and BIM. For further details, please contact Ascent Research.
