The KRT14 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HAP1 cells with targeted disruption of the KRT14 gene. This polyclonal knockout pool is generated by CRISPR/Cas9-mediated gene editing, enabling loss-of-function studies without clonal selection. Heterogeneous mutations across the population allow robust functional comparisons with wild-type controls in pooled assays.
HAP1 is a near-haploid human cell line derived from a male chronic myeloid leukemia patient, featuring an adherent fibroblast-like morphology. Its single gene copy for most loci simplifies knockout generation and reduces genetic redundancy, making it ideal for functional genomics. The myeloid background provides a distinct cellular environment for investigating keratin filament biology independent of epithelial differentiation programs.
KRT14 encodes a type I keratin that forms obligate heterodimers with KRT5, the type II keratin, to assemble intermediate filaments essential for mechanical stability in basal keratinocytes. The KRT5/KRT14 network is anchored to hemidesmosomes through plectin, BPAG1, and integrin ??6??4, while desmoplakin and plakoglobin connect filaments to desmosomes. KRT14 transcription is driven by TP63 and modulated by EGF/EGFR, Notch, and AP-1 signals. Knockout disrupts filament integrity, causing cytoskeletal disorganization, reduced cell stiffness, and compensatory upregulation of KRT15 and KRT16, ultimately compromising epithelial barrier function.
In HAP1 cells, which lack an endogenous keratin network, this knockout model provides a simplified genetic context to study KRT14 protein interactions and downstream pathways without epithelial-specific complexity. The near-haploid genome enables unambiguous analysis of binding partners and post-translational modifications. It is particularly useful for synthetic lethal screens and drug testing, as the reduced genetic background minimizes confounding compensatory mechanisms seen in diploid epithelial lines.
Applications include immunofluorescence for filament remnants, western blotting for KRT14 and KRT5, and adhesion or migration assays to assess mechanical phenotypes. Colony formation, scratch wound healing, and permeability assays evaluate collective cell behavior and barrier function when KRT14 is exogenously expressed. The cells also support drug sensitivity screens under mechanical stress, aiding in the discovery of therapies for epidermolysis bullosa simplex and other keratinopathies. For technical inquiries, please contact Ascent Research.
