The AKT2 Knockout HT29 Polyclonal Cells product consists of a heterogeneous population of human HT29 colorectal adenocarcinoma cells engineered via CRISPR/Cas9-mediated gene disruption to generate a loss-of-function model for the AKT2 gene. This polyclonal knockout pool retains the diverse genetic background of the parental cell line while eliminating functional AKT2 expression across the population, providing a versatile tool for studying AKT2-dependent biology without clonal artifacts.
HT29 cells originate from a human colorectal adenocarcinoma and exhibit epithelial morphology, representing a well-characterized model for intestinal epithelial biology and colorectal cancer research. These cells are commonly employed in investigations of tumorigenesis, differentiation, and drug response, and they harbor mutations in APC, TP53, and SMAD4, which recapitulate key oncogenic pathways in colorectal cancer.
AKT2 (also known as PKB??) encodes a serine/threonine kinase that serves as a central effector of the PI3K signaling axis. Upon activation by upstream receptor tyrosine kinases such as EGFR and IGF1R, PI3K-generated PIP3 recruits AKT2 to the plasma membrane, where it is phosphorylated by PDK1 at Thr309 and by mTORC2 at Ser474. Once activated, AKT2 phosphorylates a broad array of downstream substrates including GSK3??, TSC2, PRAS40, FOXO1, and AS160, thereby promoting cell survival, proliferation, glucose uptake, and metabolism. Its activity is tightly regulated by phosphatases like PHLPP and PP2A, as well as by interacting partners such as APPL1 and HSP90.
In the HT29 colorectal cancer background, constitutive activation of AKT2 is frequently observed and contributes to oncogenic processes including enhanced proliferation, resistance to apoptosis, and altered metabolism. Disruption of AKT2 in this polyclonal population abrogates downstream signaling through the mTORC1/S6K and FOXO pathways, thereby enabling dissection of AKT2-specific functions in colorectal tumorigenesis, insulin signaling, and chemoresistance. This model is particularly valuable for investigating isoform-specific roles of AKT2 versus AKT1 and AKT3, given the prevalent use of HT29 cells in signaling and drug discovery studies.
Researchers can employ these knockout cells in a wide range of experimental settings, including Western blotting and phospho-AKT signaling analysis to validate loss of AKT2 and its downstream targets, cell proliferation and apoptosis assays to assess AKT2-mediated survival pathways, and glucose uptake measurements to study metabolic effects. Migration and invasion assays facilitate examination of AKT2’s role in metastatic behavior, while tumor xenograft models and drug sensitivity assays allow for preclinical evaluation of therapeutic vulnerabilities. For questions about this product or custom gene editing services, please contact Ascent Research.
