The AKT2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 near-haploid human cell line, with targeted disruption of the AKT2 gene. This loss-of-function model is generated through CRISPR/Cas9-mediated gene editing, generating a heterogeneous pool of cells carrying AKT2 gene disruptions, and provides a versatile tool for studying AKT2-dependent signaling and function without the constraints of single-cell cloning.
The HAP1 cell line is a near-haploid human hematopoietic cell model derived from the KBM-7 chronic myeloid leukemia cell line, originating from a male donor. Its near-haploid karyotype simplifies genetic manipulation and phenotypic analysis, making it particularly valuable for high-throughput genetic perturbation screens and functional genomics studies. HAP1 cells retain key signaling pathways relevant to hematologic malignancies and metabolism, providing a physiologically relevant context for dissecting AKT2 biology.
AKT2 is a serine/threonine kinase central to the PI3K signaling pathway. Following insulin or growth factor stimulation, PI3K generates PIP3, which recruits AKT2 to the membrane where it is phosphorylated by PDK1 (Thr309) and mTORC2 (Ser474). Activated AKT2 phosphorylates downstream substrates such as GSK3??, FOXO transcription factors, TSC2, PRAS40, BAD, AS160, MDM2, and eNOS, driving glucose metabolism, cell survival, and proliferation. PTEN antagonizes this pathway by dephosphorylating PIP3. AKT2 also interacts with 14-3-3 proteins and HSP90, positioning it within networks regulating insulin, mTOR, FoxO, and AMPK signaling.
In the HAP1 near-haploid background, AKT2 disruption enables unhindered dissection of AKT2 isoform?specific functions without confounding compensation from paralogs. The absence of a second allele facilitates straightforward genotype?phenotype correlation, enhancing the utility of this model for loss?of?function analyses in pathways governing metabolic control and oncogenic growth. This is particularly relevant for studying insulin resistance, type 2 diabetes, and cancers such as breast, ovarian, pancreatic, and glioblastoma, where AKT2 is frequently dysregulated.
These AKT2 knockout HAP1 polyclonal cells are suitable for a range of experimental applications, including functional studies of insulin?stimulated glucose uptake, western blot analysis of AKT2 and phospho?substrates, RT?qPCR for transcript validation, and cell viability or apoptosis assays under metabolic stress or chemotherapeutic challenge. They support drug sensitivity profiling for PI3K pathway inhibitors, phospho?signaling analysis, synthetic lethality screens, and target validation in oncology and metabolic disease research. For further information, contact Ascent Research.
