The KRT18 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the KRT18 gene in the HAP1 cell line. This loss-of-function model provides a versatile tool for studying keratin 18 biology, with the polyclonal format capturing a range of editing events that collectively ablate gene function at the population level. It supports high-content screening and functional genomics without the biases of single-clone selection.
HAP1 cells are a human near-haploid fibroblast-like adherent cell line derived from the KBM-7 chronic myeloid leukemia line. The near-haploid karyotype facilitates efficient CRISPR-based knockout generation by minimizing functional redundancy from a second allele, enabling straightforward genotype-phenotype correlations. HAP1 cells retain expression of epithelial keratins, making them suitable for intermediate filament research.
KRT18 encodes keratin 18, a type I intermediate filament protein that forms obligate heteropolymers with keratin 8 (KRT8), providing mechanical stability to simple epithelium. During apoptosis, keratin 18 is cleaved by caspase-3, -6, and -7, releasing M30 fragments used as biomarkers. Keratin 18 also participates in signal transduction: it interacts with TRADD to modulate TNFR1-mediated NF-??B activation, binds 14-3-3 proteins to promote survival, and associates with Akt/PKB to sustain anti-apoptotic signaling. Expression is regulated by WNT/TCF/LEF, EGF/ERK/MAPK, and cytokines such as TNF-?? and IL-6.
In HAP1 cells, the near-haploid background simplifies analysis of KRT18 loss-of-function phenotypes. The polyclonal knockout population captures heterogeneous editing outcomes, enabling robust study of apoptosis resistance, keratin filament dynamics, and crosstalk with survival pathways. This model is particularly suited for dissecting KRT18??s role in Fas- and TNF-??-induced signaling without clonal artifacts.
Applications include studies of epithelial integrity, apoptosis, and cancer. Typical assays are western blotting for full-length and cleaved KRT18, immunofluorescence of keratin networks, flow cytometry with M30 antibody, and caspase activity assays. The cells support drug screening for modifiers of PI3K/Akt/GSK3??/??-catenin signaling and biomarker discovery. For further information, please contact Ascent Research.
