The AKT2 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with disrupted AKT2 to create a loss-of-function model. Supplied as a polyclonal pool, this product maintains genetic heterogeneity while eliminating AKT2 protein function, supporting robust analyses free of clonal selection bias.
The HCT 116 host is a human male colorectal carcinoma line with KRAS G13D and PIK3CA H1047R mutations, MSI-H status, and MLH1 deficiency, widely used for cancer biology, apoptosis, and drug sensitivity studies.
AKT2 is a serine/threonine kinase central to PI3K/AKT signaling. Activated by PIP3-mediated membrane recruitment, PDK1 phosphorylates Thr309 and mTORC2 phosphorylates Ser474. Active AKT2 phosphorylates downstream targets including GSK3?? (inactivating it to promote proliferation and glucose metabolism), FOXO1/3a (blocking pro-apoptotic transcription), BAD and Caspase-9 (inhibiting apoptosis), TSC2 (activating mTORC1), AS160 (enhancing glucose uptake), and PRAS40 (relieving mTORC1 inhibition). Negative regulators include PTEN, which dephosphorylates PIP3, and phosphatases PP2A and PHLPP that directly dephosphorylate AKT2.
Despite constitutive KRAS and PIK3CA activation in HCT 116, AKT2 knockout substantially reduces phosphorylation of key substrates like GSK3?? and FOXO, impairing proliferation and promoting apoptosis. This underscores AKT2’s indispensable role in maintaining oncogenic signaling in the context of upstream mutations, providing a unique system to study isoform-specific AKT functions.
Applications include phospho-signaling profiling by western blotting (pGSK3??, pFOXO, pPRAS40), AKT inhibitor sensitivity assays (cell viability, Annexin V/PI apoptosis), metabolic assays (glucose uptake), migration/invasion tests, and RT-qPCR for AKT2 mRNA. This polyclonal model enables studies of AKT2-dependent pathways in a clinically relevant colorectal cancer background. For technical support, contact Ascent Research.
