The KRT20 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-mediated gene-edited population of 143B osteosarcoma cells with targeted disruption of KRT20. This polyclonal pool consists of a heterogeneous mixture of edited cells carrying random allelic modifications, providing a loss-of-function model without clonal selection. Absence of KRT20 protein enables functional dissection of keratin 20 in a malignant mesenchymal background. The product is suitable for gene expression analysis, protein interaction studies, and population-level functional assays.
The 143B cell line, a thymidine kinase-negative derivative of HOS osteosarcoma, originates from a human bone tumor. Its high tumorigenicity and metastatic potential establish it as a standard model for osteosarcoma metastasis, particularly in tail vein injection pulmonary colonization assays. As a mesenchymal cell type, 143B lacks endogenous keratin expression, offering an ideal background for manipulating epithelial keratins such as KRT20. The cell line??s aggressive behavior and well-characterized genetic background provide a robust platform for investigating tumor dissemination mechanisms.
KRT20 encodes a type I intermediate filament protein that pairs with keratin KRT8 to form the epithelial cytoskeleton. Its expression is regulated by CDX2, HNF4A, GATA4, GATA6, retinoic acid, and Wnt/??-catenin signaling. The KRT20?CKRT8 filament network anchors to desmosomes via desmoplakin and plectin, while 14-3-3 scaffold proteins (YWHA) mediate downstream signal transduction. KRT20 modulates tight junction protein ZO-1 localization and is associated with vimentin upregulation and Snail transcription factor activity during epithelial-mesenchymal transition. TRADD links KRT20 to apoptotic pathways. In this knockout model, disruption of KRT20 perturbs the CDX2/KRT20 regulatory axis and desmosomal cadherin complexes, facilitating interaction mapping and functional studies.
KRT20 knockout in 143B cells creates a unique system to study keratin loss in a mesenchymal tumor context. The inherently metastatic 143B background may exhibit altered dissemination patterns upon ablation of an epithelial intermediate filament protein, potentially through cytoskeletal deregulation. This model enables investigation of cell adhesion, anoikis resistance, and extracellular matrix invasion. It also allows exploration of compensatory keratin expression and crosstalk with dysregulated pathways such as Wnt/??-catenin. The polyclonal population avoids clonal artifacts and captures heterogeneous gene perturbation responses.
Applications of this product include cancer metastasis, epithelial-mesenchymal transition, and cytoskeletal dynamics research. Standard assays encompass immunofluorescence and western blotting for KRT20 depletion confirmation, RT-qPCR for downstream target analysis, and transwell migration assays for motility assessment. Co-immunoprecipitation can map interactions with 14-3-3 proteins or desmoplakin, while in vivo tail vein metastasis assays directly evaluate pulmonary colonization. The polyclonal knockout pool serves as a starting material for anti-metastatic drug screening targeting keratin-associated adhesions. For additional technical details or customization, please contact Ascent Research.
