The HSP90AB1 Knockout DLD-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population originating from the DLD-1 human colorectal adenocarcinoma line. This product features targeted disruption of the HSP90AB1 gene, resulting in loss of HSP90?? protein function across a heterogeneous pool of edited cells. The polyclonal constitution avoids single-cell cloning, retaining the genetic diversity of the parental line while creating a robust loss-of-function model for studying HSP90?? biology in colorectal cancer. The polyclonal knockout cells are provided as a mixed population, suitable for immediate use in functional assays.
The DLD-1 cell line (ATCC CCL-221) is a widely used model of colorectal adenocarcinoma isolated from a Dukes?? type C tumor, representing an aggressive, invasive cancer stage. These epithelial cells carry oncogenic mutations in APC and KRAS, which activate Wnt and MAPK pathways, making them especially relevant for studying signal transduction and tumorigenesis. The well-defined genetic background and reliable in vitro growth support reproducible experiments investigating chaperone function in colorectal cancer.
HSP90AB1 encodes HSP90??, an ATP-dependent chaperone that stabilizes and activates client proteins including kinases (AKT, RAF), steroid receptors, HIF-1??, and survivin. Co-chaperones such as CDC37, AHA1, p23, and HOP regulate its ATPase cycle and client loading. HSP90?? is transcriptionally controlled by HSF1 and modulated by heat shock, oxidative stress, and HDAC6/SIRT1. It functions at the core of signal transduction networks such as the HSP90-CDC37-AKT, HSP90-RAF-MEK-ERK, and HSP90-HIF-1??-VEGF axes, integrating stress responses with growth and survival signaling.
In colorectal cancer, HSP90?? overexpression is common and sustains malignant phenotypes by chaperoning oncogenic drivers such as AKT and RAF. Disruption of HSP90AB1 in DLD-1 cells leads to client protein degradation, attenuates PI3K/AKT and MAPK/ERK pathways, and impairs proliferation, colony formation, and survival. This knockout model permits precise dissection of HSP90?¡?s tumorigenic role, including anchorage-independent growth and apoptosis regulation, while offering a controlled background for evaluating HSP90 inhibitor sensitivity.
Typical applications include western blotting and RT-qPCR for knockout confirmation, phospho-signaling analysis (e.g., phospho-AKT), proliferation and colony formation assays, Annexin V apoptosis detection, and co-immunoprecipitation for client identification. Drug sensitivity profiling with HSP90 inhibitors (e.g., geldanamycin) can identify vulnerabilities. The polyclonal nature is advantageous for pooled screens and heterogeneity studies. For further information, please contact Ascent Research.
