HSP90AA1 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population generated from the HCT 116 colorectal carcinoma line, targeting the HSP90AA1 gene. This product consists of a heterogeneous pool of cells, each carrying gene disruptions introduced by CRISPR/Cas9, providing a loss-of-function model without clonal selection. The polyclonal format captures diverse mutational outcomes, enabling robust assessment of functional consequences across a genetically mixed population.
The HCT 116 host cell line is a well-characterized human colorectal adenocarcinoma model featuring microsatellite instability (MSI-high), a KRAS G13D activating mutation, and wild-type p53. This genetic profile mirrors key aspects of colorectal tumorigenesis, including defective DNA mismatch repair and constitutive RAS?CMAPK signaling. HCT 116 cells are widely employed for studying oncogenic pathways, drug responses, and tumor suppressor functions in a colorectal cancer context.
HSP90AA1 encodes the stress-inducible molecular chaperone HSP90??, which facilitates the folding, stabilization, and activity of numerous client proteins, including the kinases EGFR, ERBB2, CDK4, RAF1, and AKT1, the transcription factor HIF1A, and steroid hormone receptors. HSP90AA1 expression is upregulated by HSF1 in response to heat shock, oxidative stress, and growth factor stimulation. Its chaperone cycle depends on co-chaperones such as CDC37, STIP1, PTGES3, AHSA1, and FKBP5, which regulate ATP-driven conformational changes. By maintaining client protein homeostasis, HSP90AA1 integrates signals from PI3K/AKT/mTOR, MAPK/ERK, and JAK/STAT pathways, positioning it as a critical node in proliferative and survival signaling networks.
In HCT 116 cells, knockout of HSP90AA1 disrupts chaperone function, leading to misfolding and proteasomal degradation of essential oncogenic clients. This impairs downstream proliferative and survival signaling, resulting in cell cycle arrest and induction of apoptosis. The concomitant KRAS G13D mutation sensitizes these cells to loss of chaperone activity, as mutant KRAS and other client oncoproteins rely on HSP90 for conformational maturation and signaling competence. The polyclonal knockout population thus recapitulates the cellular consequences of HSP90 inhibition in a physiologically relevant, heterogeneous cancer cell background.
This HSP90AA1 knockout model is suited for diverse research applications, including validation of HSP90 inhibitor targets, dissection of RAS?CPI3K?CAKT signaling crosstalk, and mechanistic studies of chaperone-mediated protein quality control. Compatible assays include western blotting, RT-qPCR, immunofluorescence, proliferation and apoptosis assays, cell cycle analysis, co-immunoprecipitation, and drug sensitivity testing. The polyclonal nature allows for unbiased evaluation of therapeutic vulnerabilities and chaperone dependencies. For additional technical details, please contact Ascent Research.
