The KRT5 Knockout DLD-1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed for the disruption of the KRT5 gene in the DLD-1 colorectal adenocarcinoma cell line. This product provides a genetically heterogeneous loss-of-function model, eliminating the need for single-cell cloning and enabling the study of KRT5 deficiency across a diverse allelic spectrum within a defined genetic background. The polyclonal format retains the inherent variability of a CRISPR-edited pool, offering a robust tool for functional genomics in epithelial cancer research.
The DLD-1 cell line originates from a male patient diagnosed with Dukes’ type C colorectal adenocarcinoma and serves as a well-characterized model of intestinal epithelial biology. DLD-1 cells are widely utilized in cancer research to investigate signaling pathways governing proliferation, differentiation, and metastasis, particularly in the context of adherent and invasive phenotypes. Their genetic background, including known mutations in key oncogenes and tumor suppressors, provides a relevant platform for assessing the tumorigenic impact of specific gene disruptions.
Keratin 5, encoded by KRT5, is a type II intermediate filament protein that obligatorily pairs with keratin 14 (KRT14) to form heteropolymeric filaments essential for epithelial cell structural integrity, migration, and wound healing. Upstream, KRT5 expression is transcriptionally regulated by TP63, AP-1 transcription factors, and p53, and is responsive to EGFR and TGF-?? signaling. KRT5 interacts with desmosomal components Desmoplakin, Plakoglobin, and Plakophilin, and with Integrin ??4 to anchor filaments at focal adhesions. Disruption of KRT5 in DLD-1 cells destabilizes the intermediate filament network, impairing KRT14 assembly, desmosome formation, and focal adhesion stability. This perturbation alters downstream signaling through integrin-mediated adhesion and MAPK pathways (MAPK1/3) and may modulate CTNNB1 and SRC, affecting epithelial-to-mesenchymal transition (EMT) and tumorigenic properties.
In the DLD-1 colorectal adenocarcinoma background, KRT5 knockout provides a powerful model to dissect the interplay between cytoskeletal architecture and cancer cell plasticity. The loss of keratin 5 compromises mechanical resilience and adhesive properties, making these cells particularly suited for studying EMT, collective cell migration, and drug resistance mechanisms in intestinal epithelial tumors. By disrupting pathways that link mechanical cues to transcriptional programs??via TP63 and EGFR??this knockout system enables researchers to investigate how intermediate filament dysregulation contributes to malignant progression.
This polyclonal knockout cell population is ideally suited for a broad range of experimental approaches, including immunofluorescence and cytoskeleton staining to visualize filament disorganization, scratch wound healing and Transwell migration assays to quantify migratory defects, and co-immunoprecipitation to probe protein?Cprotein interactions with KRT14 and desmosomal proteins. Western blotting and flow cytometry can confirm loss of KRT5 protein and assess EMT markers, while RNA-seq and drug sensitivity assays facilitate transcriptomic profiling and therapeutic response studies in the context of colorectal cancer. For additional details or technical inquiries, please contact Ascent Research.
