The KRT14 Knockout 786-O Polyclonal Cells product consists of a heterogeneous pool of 786-O human clear cell renal carcinoma cells subjected to CRISPR/Cas9-mediated disruption of the KRT14 gene. This polyclonal knockout population allows loss-of-function studies without single-cell clonal expansion, preserving genetic diversity while abolishing keratin 14 expression. It serves as a versatile tool for investigating KRT14-dependent processes in a cancer-relevant background.
The parental 786-O cell line is a well-characterized VHL-deficient model of clear cell renal cell carcinoma, originally established from a primary renal adenocarcinoma of a 58-year-old male. These adherent epithelial cells display constitutive stabilization of hypoxia-inducible factors, particularly HIF-1??, and are widely employed to study hypoxia-driven angiogenesis, metabolic reprogramming, and tumor progression. The line’s genetic and phenotypic features make it especially suitable for dissecting pathways that contribute to renal carcinoma aggressiveness.
Keratin 14 is a type I intermediate filament protein that obligatorily heterodimerizes with type II keratin 5 (KRT5) to form the structural backbone of epithelial intermediate filaments. KRT14 transcription is regulated by p63 and AP-1, and is further induced by TGF-??, EGF, and hypoxia through HIF-1?? and SP-1. Expressed KRT14 interacts with desmoplakin, plectin, BPAG1, and epiplakin to anchor filaments to desmosomes and hemidesmosomes. Genetic disruption of KRT14 results in loss of cytoskeletal integrity, accompanied by upregulation of mesenchymal markers such as vimentin and downregulation of epithelial markers like E-cadherin. These changes promote cell migration and epithelial-mesenchymal transition, mediated in part through altered TGF-??/SMAD2/3 signaling.
Within the VHL-null 786-O background, where HIF-1?? is constitutively active, KRT14 knockout provides a unique platform to examine the interplay between hypoxia signaling and intermediate filament dynamics. The disruption of KRT5-KRT14 heterodimers is predicted to compromise cellular tensile strength and facilitate mesenchymal conversion, thus modeling aspects of sarcomatoid differentiation observed in aggressive renal cell carcinoma. This system enables functional dissection of keratin-dependent cytoskeletal remodeling under normoxic and hypoxic conditions.
The polyclonal knockout cells are suited for a broad range of applications, including investigation of epithelial-mesenchymal transition, metastasis, cytoskeletal reorganization during hypoxia, and drug resistance mechanisms linked to keratin expression. Representative assays include Western blotting for EMT markers (E-cadherin, vimentin), immunofluorescence to visualize intermediate filament networks, Boyden chamber and wound-healing migration/invasion assays, co-immunoprecipitation to assess KRT5-KRT14 complex formation, and RNA-seq for global transcriptomic profiling. For further information, please contact Ascent Research.
