The EHF Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the haploid human HAP1 cell line, engineered to disrupt the ETS homologous factor (EHF) gene. This polyclonal pool provides a heterogeneous loss-of-function model, enabling robust functional studies without clonal isolation. The product is supplied as a mixed population carrying diverse Cas9-induced mutations, suitable for pooled screening and averaging phenotypic variability.
HAP1 cells originate from the near-haploid KBM-7 chronic myeloid leukemia line, maintained as a haploid model for genetic screens. Their haploid genome simplifies knockout generation, as a single mutation suffices for complete gene disruption. Although non-epithelial, HAP1 cells serve as a clean background to dissect transcription factors like EHF that govern epithelial identity, offering a standardized platform for functional genomics and drug target validation.
EHF is an ETS family transcription factor driving epithelial differentiation and barrier maintenance. It is activated by EGFR ligands (EGF, TGF-alpha) and TGF-beta, relaying signals through the RAS-RAF-MEK-ERK and TGFBR1-SMAD2/3-SMAD4 pathways. Together with coactivators SPDEF, FOXA2, and p300, EHF transcriptionally upregulates tight junction components such as CLDN1, mucins like MUC5AC, and E-cadherin, while repressing mesenchymal programs. Additionally, EHF promotes proliferation via Cyclin D1. Thus, EHF integrates multiple signals to orchestrate epithelial architecture and mucosal integrity.
In the HAP1 background, EHF knockout disrupts key epithelial gene programs, even in non-epithelial cells, providing a tractable system to study barrier dysfunction. The polyclonal format minimizes clonal effects, making it ideal for examining diseases like colorectal cancer, lung adenocarcinoma, asthma, and inflammatory bowel disease, where EHF loss impairs tight junctions and differentiation. This model enables dissection of EHF-dependent mechanisms in a haploid, easily manipulable context.
This product supports diverse applications: drug screens for epithelial barrier modulators, functional genomics of transcriptional regulation, and pathway analysis of EHF targets. Validation strategies include Western blot for EHF, CLDN1, E-cadherin; RT-qPCR for MUC5AC, SPDEF; RNA-seq for global transcriptomics; and immunofluorescence for junctional markers. Functional assays like TEER and migration assays can be adapted following ectopic expression of relevant adhesion molecules, with flow cytometry quantifying surface epithelial markers. For ordering and technical support, contact Ascent Research.
