The KRT77 Knockout HAP1 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the KRT77 gene in the HAP1 host cell line. This knockout model enables loss-of-function studies of KRT77, a type II keratin that functions as a structural component of keratin intermediate filaments in suprabasal epithelial layers. The polyclonal format supports robust experimental analysis without single-cell cloning artifacts, making it suitable for population-level signaling and structural investigations.
HAP1 cells are a near-haploid human male cell line derived from the KBM-7 chronic myeloid leukemia (CML) line, which is Philadelphia chromosome?Cpositive and isolated from a patient in blast crisis. This host line retains a CML disease background while offering simplified genetic analysis due to its haploid state. Widely used for functional genomics and knockout screens, HAP1 cells provide a streamlined platform for evaluating gene function and drug sensitivity in a leukemia context.
KRT77 contributes to mechanical stability and barrier integrity in epithelial tissues through its incorporation into keratin intermediate filament networks. It interacts with filament partners including KRT18, KRT5, and KRT14, and with desmosomal and adhesion complex components such as desmoplakin, plakoglobin, BPAG1e, and epiplakin. Upstream regulation of KRT77 involves EGFR ligands, calcium influx, retinoic acid receptors, AP-1 transcription factors (c-Fos/c-Jun), p63/p73, C/EBP??/??, Notch intracellular domain, and TGF-??/Smad signaling. Downstream consequences of KRT77 loss include altered keratin filament architecture, reduced cellular mechanical stiffness, impaired desmosome assembly, and dysregulated MAPK/ERK signaling, potentially through altered EGFR trafficking. The associated network converges on pathways like keratinocyte differentiation, intermediate filament organization, EGFR signaling, Notch signaling, hemidesmosome assembly, focal adhesion dynamics, and epithelial-to-mesenchymal transition.
In the HAP1 background, KRT77 knockout disrupts the keratin cytoskeleton, compromising tensile strength and cell adhesion. This perturbation alters EGFR trafficking and downstream MAPK/ERK and PI3K/AKT pathways, promoting migratory and proliferative phenotypes. The CML origin of HAP1 cells further allows investigation into how intermediate filament remodeling influences oncogenic signaling and imatinib sensitivity, providing insights relevant to drug resistance mechanisms.
This polyclonal knockout population is suited for dissecting keratin network mechanics, epithelial barrier function studies, screening for keratinopathy therapeutics, and investigating cross-talk between intermediate filaments and signaling pathways. Typical assays include western blotting, RT-qPCR, immunofluorescence for keratin filaments, scratch wound migration, transwell invasion, co-immunoprecipitation of keratin complexes, phospho-ERK1/2 flow cytometry, imatinib sensitivity dose?Cresponse, and RNA-seq differential expression analysis. For further product details or technical support, contact Ascent Research.
