The AKT1 Knockout DLD-1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population, providing a loss-of-function model for the AKT1 serine/threonine kinase. These polyclonal knockouts arise from gene disruption via CRISPR/Cas9, avoiding clonal artifacts and enabling population-level analyses. The model is designed for investigating AKT1-dependent cellular processes in colorectal cancer, circumventing pharmacological inhibitor limitations.
DLD-1 is a widely used human colorectal adenocarcinoma cell line derived from a Dukes’ type C tumor. It exhibits invasive properties and carries mutations in APC, KRAS, and TP53, reflecting advanced colorectal cancer. The line’s adherent growth and well-defined signaling pathways make it suitable for standardized assays of proliferation, apoptosis, and drug response, which are strongly influenced by PI3K/AKT activity.
AKT1 operates as a central kinase in the PI3K/AKT cascade, activated downstream of receptor tyrosine kinases (EGFR, PDGFR, insulin receptor) via PI3K, PDK1, and mTORC2. Once active, AKT1 phosphorylates substrates such as mTOR, GSK3??, BAD, FoxO1, and TSC2, driving survival, proliferation, and metabolic reprogramming. PTEN and PP2A act as negative regulators, while HSP90 stabilizes AKT1. This axis is frequently dysregulated in colorectal cancer through PIK3CA mutations or PTEN loss.
In DLD-1 cells, AKT1 hyperactivity promotes anchorage-independent growth, metabolic shifts, and resistance to apoptosis. CRISPR/Cas9-mediated knockout of AKT1 disrupts these oncogenic signals, creating a clean genetic background to dissect AKT1-specific roles and potential compensation by AKT2/3. This model is particularly useful for studying colorectal cancer signaling addiction and resistance to PI3K/mTOR inhibitors, with the polyclonal format minimizing single-cell biases.
These cells support diverse assays, including viability assessments (MTT/CellTiter-Glo), apoptosis analysis (flow cytometry with Annexin V), and migration/invasion studies (Transwell). They are also employed in metabolic profiling, drug combination screens, and mechanistic work monitoring phospho-AKT by western blotting or expression of targets like FoxO1 by RT-qPCR. Protein interactions, for example with HSP90, can be examined by co-immunoprecipitation. For further technical details, please contact Ascent Research.
