The HSPB1 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the HSPB1 (Hsp27) gene, generating a loss-of-function model. This heterogeneous population, derived without single-cell cloning, minimizes clonal artifacts and provides a robust platform for functional studies.
The parental HCT 116 cell line is a human colorectal carcinoma epithelial model characterized by tumorigenicity and invasiveness. It harbors a KRAS G13D mutation, ??-catenin S45 deletion, and MLH1 deficiency leading to microsatellite instability-high (MSI-H), which drive dysregulated proliferation, Wnt/??-catenin signaling, and defective DNA mismatch repair, making it a relevant system for colon cancer research.
HSPB27 functions as an ATP-independent molecular chaperone and anti-apoptotic regulator. Under stress, p38 MAPK activates MAPKAPK2, which phosphorylates HSPB1 at Ser15, Ser78, and Ser82, promoting oligomer-to-dimer transition. Phospho-HSPB1 dimers bind and inhibit cytochrome c and procaspase-3, preventing apoptosome formation and caspase-9/3 activation. HSPB1 also stabilizes F-actin, activates AKT, and may modulate NF-??B via I??B?? interaction, collectively enhancing cell survival, migration, and chemoresistance.
In HCT 116 cells, HSPB1 knockout ablates the cytoprotective response to genotoxic and oncogenic stress, sensitizing cells to chemotherapeutics like 5-fluorouracil and oxaliplatin. This model enables dissection of drug resistance mechanisms and apoptosis regulation in a genetically defined colorectal cancer background, reflecting the heterogeneity of tumor cell populations.
Typical applications include Western blotting and RT-qPCR for HSPB1 expression, immunofluorescence for actin cytoskeleton analysis, flow cytometry for apoptosis (Annexin V/PI), and caspase activity assays. It also supports migration/invasion studies (wound healing, Transwell), co-immunoprecipitation for protein interactions, phospho-signaling analysis, and drug sensitivity profiling. For further technical support, contact Ascent Research.
