The KRT8 Knouckout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population for loss-of-function studies of human KRT8. This heterogeneous pool of HAP1 cells harbors targeted disruptions in the KRT8 locus, generated without single-cell cloning. The polyclonal format maintains genetic diversity, enabling robust functional readouts and pooled screening approaches. Researchers can exploit this near-haploid knockout model to dissect KRT8-dependent processes with minimal genetic compensation.
HAP1 is a near-haploid human myeloid cell line derived from KBM-7 chronic myeloid leukemia cells, exhibiting fibroblast-like adhesive morphology and a predominantly haploid karyotype. This genetic simplicity accelerates complete loss-of-function allele generation and is widely used in chemogenomic screens and genetic interaction mapping. Its haploid nature reduces confounding variability, making it a gold standard for studying gene essentiality and drug mechanisms.
KRT8 encodes a type II keratin that pairs with KRT18 to form intermediate filaments in simple epithelia. Beyond structural support, KRT8 dynamically integrates mechanical stress, survival, and apoptosis signals. Phosphorylation, stimulated by EGFR, TNF-??, and IL-1??, recruits adaptor proteins like 14-3-3, plectin, desmoplakin, and HRAS, modulating FAK, Akt, and Erk. Thus, KRT8 sits at the nexus of TNF and PI3K/Akt pathways, translating extracellular cues to cytoskeletal reorganization, cell adhesion dynamics, and transcriptional outputs via GATA4 and p63.
In the HAP1 myeloid background, KRT8 disruption uncovers non-canonical roles of intermediate filaments. Haploid genetics ensure that phenotypic effects directly reflect KRT8 loss, providing an ideal system for studying apoptosis and drug resistance mechanisms pertinent to myeloid leukemia. As a downstream effector of TNF-?? and PI3K/Akt??pathways often aberrant in hematopoietic cancers??this model reveals crosstalk between keratin dynamics and cell death.
This polyclonal knockout facilitates diverse applications. Immunoblotting for KRT8/KRT18 and immunofluorescence microscopy confirm knockout and filament collapse. Cell migration and adhesion assays address integrin-mediated mechanotransduction. Apoptosis (Annexin V/PI) and phospho-proteomics uncover signaling changes downstream of EGFR or TNF-??. Drug sensitivity screens enable identification of synthetic lethalities, particularly relevant to hepatocellular carcinoma and colorectal cancer. For further details, contact Ascent Research.
