The EGF Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population in which the EGF gene has been disrupted to create a loss-of-function model. This heterogeneous pool of edited cells reduces clonal artifacts and is well suited for population-level functional studies, including pooled screening assays.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) cell line. It exhibits a stable, adherent morphology and retains a predominantly haploid karyotype, greatly simplifying genetic manipulation. Because most genes exist in a single copy, knockout of EGF yields a near-complete loss of gene function without compensatory effects from a second allele, making HAP1 an advantageous platform for dissecting signaling pathways that are otherwise confounded by diploidy.
EGF encodes a secreted growth factor that binds the epidermal growth factor receptor (EGFR/ERBB1), inducing receptor dimerization and autophosphorylation. This recruits the adaptor protein GRB2, leading to activation of the RAS-RAF-MEK-ERK (MAPK) cascade and transcriptional induction of downstream targets including MYC, FOS, JUN, and CCND1. Concurrently, the PI3K-AKT-mTOR pathway promotes cell survival and growth, with additional input from PLC??-PKC and JAK-STAT branches. EGF expression is regulated by ADAM17-mediated ectodomain shedding and transcriptional activators such as SP1, and its secretion is stimulated by hormonal and cytokine signals.
In the HAP1 background, disruption of EGF eliminates the principal ligand for EGFR, thereby abrogating autocrine and paracrine activation of downstream MAPK/ERK and PI3K/AKT cascades. The near-haploid state ensures a definitive null phenotype, enabling clear attribution of signaling outcomes to EGF-EGFR engagement. This knockout is particularly relevant for cancer research, as dysregulated EGF/EGFR signaling drives uncontrolled proliferation in many tumors. Moreover, the HAP1 CML origin allows investigation of growth factor dependencies in leukemic contexts, potentially revealing vulnerabilities in hematopoietic malignancies.
Typical applications include western blotting and RT-qPCR for monitoring pathway activity, co-immunoprecipitation to assess EGFR complex formation, and functional assays such as MTT proliferation, migration, and drug sensitivity screens targeting EGFR or downstream kinases (e.g., MEK, AKT). The polyclonal nature supports pooled CRISPR screens and long-term culture without clonal drift. The cells are also useful for validating anti-EGF antibody specificity and for testing efficacy of EGFR inhibitors. For technical inquiries, please contact Ascent Research.
