KRT78 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for functional investigation of keratin-dependent processes in a near-haploid human genetic background. This product disrupts the KRT78 gene in the HAP1 cell line, creating a loss-of-function model that enables researchers to dissect the role of the type II keratin KRT78 without the confounding effects of a diploid genome. The polyclonal format reflects a mixed population of edited cells, providing a robust and versatile tool for high-throughput screening and mechanistic studies where clonal homogeneity is not required. Typical readouts include immunoblotting, immunofluorescence, and phenotypic assays that evaluate cytoskeletal integrity and cell behavior. Researchers can use this model to generate reproducible data on keratin network disruption in a cell line optimized for genetic tractability.
The host HAP1 cell line is a human near-haploid line originally derived from the KBM-7 chronic myeloid leukemia (CML) clone, exhibiting a largely haploid karyotype except for a disomic region on chromosome 15. This unique genetic constitution simplifies knockout engineering, as only one allele typically requires targeting to achieve functional gene disruption, and facilitates unambiguous genotype-phenotype correlation in loss-of-function screens. HAP1 cells retain many features of the parental CML lineage while offering broad utility in genetic, pharmacological, and cell biological studies. Their adherent growth and robust proliferation in standard culture conditions make them suitable for a wide range of experimental workflows, from arrayed CRISPR screens to live-cell imaging of cytoskeletal dynamics.
KRT78 encodes a type II keratin intermediate filament protein that co-assembles with type I keratins such as KRT18 to form heteropolymeric filaments integral to the epithelial cytoskeleton. Its expression is regulated by the transcription factor p63 and AP-1 family members, and it participates in signaling downstream of the epidermal growth factor receptor (EGFR). KRT78-containing filaments connect to desmosomal components and cell adhesion complexes via interacting factors like plakins and adaptor proteins, thereby linking the intermediate filament network to actin microfilaments, integrins, and focal adhesions. This molecular framework underpins the mechanical resilience, shape maintenance, and migratory capacity of epithelial cells. Loss of KRT78 disrupts the assembly and organization of the keratin filament network, compromising cellular adhesion and directed migration.
In the HAP1 context, KRT78 knockout provides a simplified model to study keratin biology without the functional redundancy often present in diploid cells. The near-haploid background amplifies phenotypic consequences of the disruption, enhancing the ability to detect subtle defects in cell adhesion, spreading, and migration. This system is particularly advantageous for genetic interaction screens, where the single-copy genomic landscape reduces complexity and improves the identification of synthetic lethal or modifier relationships. Moreover, the CML-derived background retains epithelial characteristics and deregulated kinase signaling, offering a relevant platform for investigating how keratin network perturbations intersect with oncogenic pathways, including EGFR-driven signaling, and influence drug sensitivity.
Typical research applications include functional genomics, keratin cell biology, cancer cell migration studies, genetic interaction screens, and drug target validation. Experimentally, users can assess KRT78 protein loss by western blotting, visualize filament collapse and redistribution by immunofluorescence microscopy, and quantify functional effects using migration/invasion assays (e.g., transwell or scratch wound assays), cell adhesion assays on defined matrices, and drug sensitivity profiling. This polyclonal population is well-suited for pooled CRISPR screens and large-scale phenotypic analyses where the average knockout effect in a mixed population is informative. For further technical details or custom inquiries, please contact Ascent Research.
