EFNB1 Knockout HAP1 Polyclonal Cells are a pool of CRISPR/Cas9-edited HAP1 cells carrying targeted gene disruption at the EFNB1 locus, generating a polyclonal knockout population. This format avoids clonal selection bias and ensures a representative knockout phenotype, enabling robust loss-of-function studies. By eliminating ephrin-B1 expression, this model allows researchers to investigate its roles in Eph receptor signaling and cellular functions without residual gene activity.
HAP1 cells are a near-haploid human cell line derived from a patient with chronic myelogenous leukemia (CML). Their haploid karyotype facilitates complete gene knockout with a single editing event and makes them ideal for haploid genetic screens, drug sensitivity assays, and reverse genetics. The leukemic background also provides a relevant context for studying cancer signaling pathways, including Eph-ephrin networks that are often altered in hematological malignancies.
The EFNB1 gene encodes ephrin-B1, a transmembrane ligand that binds Eph receptor tyrosine kinases, primarily EphA4, EphB2, and EphB3, to initiate bidirectional signaling. Reverse signaling through ephrin-B1??s intracellular domain recruits Src family kinases and PDZ proteins such as GRIP1 and syntenin, activating FAK and p130Cas and regulating Rho GTPases (RhoA, Rac1) to control cytoskeletal dynamics. These events feed into MAPK/ERK and JNK cascades, modulating cell adhesion and migration. EFNB1 transcription is governed by HOX factors, TGF-??, and FGF signals, integrating ephrin-B1 into developmental and oncogenic programs.
In the HAP1 background, EFNB1 disruption creates a powerful model to dissect ephrin-B1 functions in a genetically simple leukemic context. The haploid state simplifies genetic interaction mapping and synthetic lethal screens to identify ephrin-B1 modifiers. Loss of ephrin-B1 impairs Eph receptor-mediated signaling that governs cell migration, invasion, and boundary formation, making this model valuable for studying craniofrontonasal syndrome, cancer metastasis, and neural development disorders where EFNB1 mutations or dysregulation occur.
These polyclonal knockout cells are suited for Transwell migration assays, cell adhesion assays on Eph receptor substrates, Western blot analysis of phosphorylated Eph receptors, FAK, ERK, and JNK, and immunofluorescence for F-actin. Co-immunoprecipitation can probe ephrin-B1 complex formation with Eph receptors and PDZ scaffolds. They also enable haploid genetic screens to uncover novel signaling modulators and drug response studies targeting Eph-ephrin pathways. For additional details or a quote, please contact Ascent Research.
