The AKT3 Knockout 143B Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the AKT3 gene has been disrupted to generate a loss-of-function model. This product provides a heterogeneous pool of 143B osteosarcoma cells carrying diverse editing events, enabling the study of AKT3 deficiency within a genetically variable cellular context. The polyclonal format is well suited for pooled functional screens, bulk signaling analyses, and pathway dissection where clonal representation of the knockout phenotype is desired.
The 143B cell line is a human osteosarcoma model derived from TE85, exhibiting an osteoblast-like phenotype with high metastatic potential. Widely utilized for metastasis and bone cancer research, 143B cells display aggressive in vivo behavior, making them a stringent system for investigating tumor progression and metastatic dissemination. Their robust growth in culture and well-characterized genetic background support rigorous examination of oncogenic signaling cascades.
AKT3 encodes a serine/threonine kinase that functions as a critical effector of PI3K signaling. Upon stimulation of growth factor receptors such as EGFR, PDGFR, and the insulin receptor, PI3K generates PIP3, which recruits AKT3 to the plasma membrane. Activation requires phosphorylation at Thr308 by PDK1 and Ser473 by mTORC2. Active AKT3 phosphorylates downstream substrates including GSK3??, FOXO1/3, BAD, MDM2, AS160, and eNOS, thereby promoting cell survival, growth, proliferation, metabolism, and angiogenesis while suppressing apoptosis. Interacting partners like HSP90, CTMP, PP2A, and PHLPP fine-tune its activity and signal duration.
In the 143B osteosarcoma context, AKT3 knockout enables dissection of PI3K/AKT-dependent mechanisms underlying the aggressive and metastatic properties of this cell line. Disruption of AKT3 in these cells allows researchers to assess its contribution to anchorage-independent growth, invasion, and resistance to apoptosis, processes frequently dysregulated in osteosarcoma. Moreover, the model facilitates the study of compensatory signaling from other AKT isoforms and cross-talk with parallel oncogenic pathways, providing insights into therapeutic vulnerabilities and resistance mechanisms.
Research applications span cancer biology, signaling, and drug development. The polyclonal population is ideal for screening AKT inhibitor sensitivity, performing phospho-signaling arrays, and conducting migration/invasion assays. It can be used in RNA-seq or proteomic analyses to uncover AKT3-dependent transcriptional networks, as well as in FOXO reporter assays. Additional applications include co-immunoprecipitation studies and glucose uptake assays. For further information, please contact Ascent Research.
