The KRT18 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with targeted disruption of the KRT18 gene in the human 143B osteosarcoma line. This loss-of-function model allows investigation of keratin 18 biology without clonal selection bias, maintaining heterogeneous genetic background while achieving robust gene inactivation. Supplied as a polyclonal pool, the cells are ready for functional studies and side-by-side comparisons with wild-type controls.
The 143B cell line, derived from a primary osteosarcoma, is a widely used model for tumor aggressiveness and metastasis. These cells display rapid growth, invasiveness, and osteolytic lesion formation in vivo, providing a stringent platform to study genes involved in metastatic progression. The mesenchymal context of 143B enables interrogation of epithelial-associated factors like KRT18 in a tumor-relevant setting.
KRT18 encodes a type I intermediate filament protein that obligatorily heterodimerizes with keratin 8 (KRT8) to form the cytoplasmic network providing mechanical resilience to epithelial cells. During apoptosis, KRT18 is cleaved by caspases, releasing signature fragments into circulation that serve as diagnostic indicators of cell death. Its expression is transcriptionally controlled by Wnt/??-catenin, EGF, retinoic acid, and the transcription factors SP1 and AP-1. Keratin 18 interacts directly with 14-3-3 proteins, TRADD, and the Fas receptor, and modulates Akt signaling, coupling structural functions to survival and death pathways. Key apoptosis regulators, including CASP3, CASP9, BCL2, and FAS, are functionally linked to KRT18-mediated signaling.
Knockout of KRT18 in 143B cells disrupts its potential modulation of apoptosis and cytoskeletal integrity, offering a tool to dissect epithelial protein contributions within a mesenchymal tumor environment. This model may uncover roles in epithelial-mesenchymal transition, chemoresistance, and metastatic dissemination. Given the established use of KRT18 cleavage fragments as non-invasive biomarkers in hepatocellular carcinoma, breast cancer, and liver disease, this system enables validation of KRT18-dependent death pathways and preclinical biomarker studies.
Applications include Western blotting and immunofluorescence for protein expression analysis, Annexin V/PI staining and caspase activity assays for apoptosis profiling, and RT-qPCR for transcriptional assessment. Migration and invasion assays, in combination with 3D culture, allow study of KRT18-dependent metastatic properties. This polyclonal knockout pool provides a versatile resource for osteosarcoma research, apoptosis studies, and biomarker development. For further details, contact Ascent Research.
