The AKT3 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-mediated polyclonal knockout cell population designed for the disruption of the human AKT3 gene. This gene-edited product provides a pooled heterogeneous population of HAP1 cells carrying loss-of-function modifications in the AKT3 locus, enabling robust functional studies without clonal selection. The polyclonal format retains genetic diversity, reducing artifacts associated with single-cell cloning and better representing population-level responses. This model serves as a versatile tool for investigating AKT3-dependent signaling pathways and cellular processes in a near-haploid human background.
HAP1 cells, derived from the KBM-7 chronic myeloid leukemia line, are a male, near-haploid human cell line with an adherent, fibroblast-like morphology. Their haploid karyotype (except for a disomic chromosome 8) facilitates loss-of-function studies, as disruption of a single allele typically yields phenotypic effects. This line is extensively used in functional genomics and genetic screens, offering a clean, isogenic background for investigating gene function, particularly in cancer-relevant pathways.
AKT3 encodes a serine/threonine kinase that is a central effector of the PI3K/AKT pathway. Activated by ligands such as IGF-1, EGF, and insulin through receptors like IGF1R and EGFR, PI3K generates PIP3, which recruits AKT3 to the membrane. PDK1 and mTORC2 then phosphorylate and activate AKT3, while PTEN counteracts this by dephosphorylating PIP3. Active AKT3 phosphorylates substrates including GSK3??, TSC2, PRAS40, FOXO1/3, MDM2, and BAD, thereby driving cell survival, proliferation, metabolism, and angiogenesis, and inhibiting apoptosis. AKT3 also promotes mTORC1 activity via TSC2 inhibition. Regulatory interactors such as HSP90, PP2A, PHLPP, CTMP, and APPL1 fine-tune AKT3 function.
The near-haploid HAP1 background offers a simplified genetic system for dissecting AKT3 isoform-specific functions. Disruption of the single AKT3 allele likely yields complete protein loss, enabling clear phenotypes. This model allows differentiation of AKT3 roles from AKT1/AKT2 in survival, metabolism, and transformation. The polyclonal AKT3 knockout population is particularly valuable for studying signaling rewiring and compensation upon AKT3 loss, relevant to melanoma, glioblastoma, and overgrowth syndromes.
Key applications include functional genomics screens for synthetic lethality, cancer biology studies of AKT3-dependent growth, and drug resistance research in melanoma and glioblastoma. Assays such as proliferation, apoptosis, migration, and immunofluorescence, along with western blotting for phospho-AKT and phospho-GSK3??, are commonly employed. The polyclonal nature supports high-throughput drug sensitivity screens. For further information, contact Ascent Research.
