The AKT3 Knockout DLD-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal adenocarcinoma DLD-1 cell line. This product features targeted AKT3 gene disruption via CRISPR/Cas9, generating a heterogeneous loss-of-function pool that better represents tumor cell diversity than clonal lines, making it ideal for studying AKT3-dependent signaling in colorectal cancer.
The DLD-1 cell line, established from a colorectal adenocarcinoma of a male patient, is widely utilized in colorectal cancer research. It harbors homozygous APC and heterozygous TP53 mutations, leading to constitutive Wnt/??-catenin pathway activation and impaired p53 function, respectively. These genetic alterations make DLD-1 a relevant model for studying tumorigenesis, and its adherent epithelial morphology facilitates standard in vitro assays and genetic manipulation.
AKT3 is a serine/threonine kinase that acts as a central node in the PI3K-AKT pathway. Upon growth factor stimulation, PI3K generates PIP3 at the membrane, recruiting AKT3 via its PH domain. PDK1 and mTORC2 then phosphorylate AKT3 at Thr305 and Ser472, respectively, fully activating the kinase. Active AKT3 phosphorylates multiple substrates, including mTORC1 (via TSC2 and PRAS40), GSK3??, FOXO transcription factors, and the pro-apoptotic protein BAD, thereby promoting protein synthesis, proliferation, glucose metabolism, and survival while suppressing apoptosis. The pathway is antagonized by the lipid phosphatase PTEN. AKT3 also interacts with 14-3-3 proteins and HSP90, which modulate its activity and stability. Aberrant AKT3 signaling contributes to the pathogenesis of glioblastoma, melanoma, and colorectal cancer.
In DLD-1 cells with pre-existing APC and TP53 mutations, AKT3 knockout provides a defined system to study AKT isoform-specific contributions to colorectal cancer. It enables investigation of PI3K pathway output under constitutive Wnt activation and compromised p53, and helps determine whether AKT3 is required for malignant traits such as anchorage-independent growth, migration, and apoptosis resistance. This model also supports analysis of compensatory signaling among AKT family members and the effects of pharmacological PI3K pathway inhibition.
This cell model is suitable for proliferation (MTT, BrdU), apoptosis (Annexin V/PI), colony formation, and migration/invasion (Boyden chamber) assays. It can be used in western blotting and RT-qPCR for AKT3 and downstream target analysis, and in drug sensitivity screens with PI3K/AKT/mTOR inhibitors. Additional applications include co-culture, 3D organoids, and xenograft studies. For further information, please contact Ascent Research.
