AKT1 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout population targeting the AKT1 gene in the HAP1 cell line. This loss-of-function model enables investigation of AKT1-dependent signaling without specifying precise editing outcomes, offering a polyclonal pool for robust functional studies. The product is ideal for researchers studying PI3K/AKT/mTOR pathway dynamics, cancer cell biology, and metabolic regulation.
HAP1 cells are a near-haploid human cell line derived from the male chronic myeloid leukemia line KBM-7. Their haploid nature facilitates gene editing and genetic screens, as a single targeting event can disrupt gene function. They retain key signaling pathways relevant to cancer research and are widely employed in functional genomics to identify drug targets.
AKT1 encodes a serine/threonine kinase central to the PI3K/AKT/mTOR axis. Upon growth factor stimulation, PI3K generates PIP3, recruiting AKT1 to the membrane where PDK1 and mTORC2 phosphorylate Thr308 and Ser473, respectively. Activated AKT1 phosphorylates numerous substrates, including GSK-3??, TSC2, Bad, and FoxO transcription factors, promoting cell survival, proliferation, and metabolism. AKT1 also interacts with HSP90, PP2A, and 14-3-3 proteins, and its activity is counteracted by PTEN. This kinase is a key node in insulin signaling and apoptotic regulation, with dysregulation linked to cancers such as breast, prostate, and glioblastoma. AKT1-mediated phosphorylation of TSC2 relieves inhibition of mTORC1, driving protein synthesis and cell growth.
In the HAP1 background, AKT1 disruption allows dissection of these pathways in a simplified genetic context. The near-haploidy reduces genetic redundancy, making phenotypic effects of knockout more pronounced. This knockout model thus enables precise dissection of AKT1??s role in proliferation, survival, and metabolic reprogramming in a cancer-relevant genetic background. It is valuable for studying AKT1??s function in leukemia signaling, drug response, and cross-talk with MAPK/ERK pathways.
Researchers can employ these polyclonal cells in various assays including Western blotting for phospho-AKT and downstream targets, cell viability assays (MTT, CellTiter-Glo), apoptosis detection via Annexin V/PI staining, cell cycle analysis by flow cytometry, and RT-qPCR for AKT1 mRNA levels. They are suitable for high-throughput screening of AKT inhibitors such as MK-2206 or Ipatasertib, and phospho-kinase arrays. These polyclonal knockout cells are provided as a ready-to-use population for immediate culture and experimentation, eliminating the need for clonal isolation. For further details, please contact Ascent Research.
