The HSF1 Knockout HT29 Polyclonal Cells are a pool of CRISPR/Cas9-edited human HT29 colorectal adenocarcinoma cells carrying targeted disruptions in the HSF1 gene. This polyclonal knockout population is generated without single-cell cloning, preserving the genetic heterogeneity inherent to the bulk edited population and enabling robust loss-of-function analyses in a cancer cell context. The product offers a convenient tool to study HSF1-dependent pathways critical for stress responses and oncogenesis, without relying on clonal artifacts. It is suitable for academic and pharmaceutical research requiring a genetically defined human colorectal cancer model with HSF1 deficiency.
The HT29 cell line originates from a human colorectal adenocarcinoma and exhibits epithelial morphology with the ability to produce mucin and undergo enterocytic differentiation. These cells harbor a homozygous R273H mutation in the TP53 tumor suppressor gene and an inactivating mutation in the APC gene, both of which are frequent events in colorectal carcinogenesis. HT29 cells serve as a well-established intestinal epithelial model for investigating colorectal cancer biology, including proliferation, apoptosis, and therapeutic responses.
HSF1 encodes the master transcriptional regulator of the heat shock response, orchestrating cellular proteostasis. Upon activation by stress or oncogenic signals, HSF1 trimerizes, binds heat shock elements, and induces chaperones such as HSP27, HSP70, and HSP90, along with anti-apoptotic factors Bcl-2 and Survivin. Its activity is governed by MAPK/ERK1/2 and AKT-mediated phosphorylation, SIRT1 deacetylation, and HSP90-mediated negative regulation, while mTORC1 promotes its translation. HSF1 also interacts with transcriptional regulators including STAT1, NF-??B, and BRG1. In cancer, HSF1 drives a transcriptional program supporting proliferation, survival, and metastasis independently of heat shock, in part by modulating p53, cyclin D1, and LKB1.
In the HT29 colorectal adenocarcinoma background, with mutant p53 and APC, HSF1 knockout allows dissection of its contribution to oncogenic processes such as apoptosis resistance, proteotoxic stress adaptation, and drug sensitivity. As HSF1 is overexpressed in colorectal cancers and linked to poor prognosis, this polyclonal model provides a relevant system to study HSF1-dependent malignant phenotypes, including interactions with mutant p53 gain-of-function and APC-loss signaling. It is ideal for investigating the heat shock response in colorectal cancer and evaluating HSF1 as a therapeutic target.
These HSF1 knockout cells are suited for diverse assays: heat shock treatment followed by Western blot for HSF1 and targets, RT-qPCR, and HSE-luciferase reporter assays. Functional studies include MTT viability, clonogenic survival, Annexin V apoptosis, proteasome activity, transwell migration, and 5-FU chemosensitivity testing. Co-immunoprecipitation and immunofluorescence enable analysis of protein interactions and localization. Please contact Ascent Research for further details.
