The EFNB2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid HAP1 human cell line, with targeted disruption of the EFNB2 gene encoding ephrin-B2. The polyclonal format comprises a heterogeneous pool of edited cells carrying diverse gene-disrupting mutations, enabling loss-of-function studies without clonal isolation. This product facilitates rapid functional genomics applications focused on ephrin-B2?Cdependent phenotypes.
HAP1 is a near-haploid fibroblast-like cell line derived from the KBM-7 chronic myeloid leukemia line, featuring a haploid chromosome set except for a disomy of chromosome 8. This genetic simplicity enhances the penetrance of single-allele knockouts, making HAP1 an ideal host for CRISPR-based gene disruption and high-throughput genetic screens due to its robust growth and adhesion properties.
EFNB2 encodes ephrin-B2, a transmembrane ligand for Eph receptors, notably EphB4 and EphA3. Ephrin-B2?CEph interactions mediate bidirectional signaling: forward signaling via the receptor activates Src kinases, FAK, and p120 RasGAP to regulate adhesion and cytoskeletal dynamics, while reverse signaling through ephrin-B2 recruits adaptors such as Grb4 (Nck2) and PDZ proteins (Pick1, syntenin) to link to MAPK/ERK, PI3K-Akt, and RhoA/ROCK cascades. Upstream regulators include Wnt/??-catenin, TGF-??, and HIF-1??, and downstream effectors include Erk1/2, Akt, and Rac1, positioning EFNB2 as a key node in migration, axon guidance, and angiogenesis.
In the HAP1 background, EFNB2 knockout allows clear dissection of ephrin-B2?Cdependent signaling without allelic redundancy, as the haploid genome ensures strong loss-of-function phenotypes. The fibroblast-like morphology and expression of Eph/ephrin components make these cells particularly suitable for studying bidirectional signaling, integrin crosstalk, and cell migration mechanisms. Given the role of ephrin-B2 in cancer and vascular biology, this model is valuable for investigating adhesion, invasion, and survival pathways relevant to glioblastoma, gastric, and breast cancers.
Applications include Western blotting for EFNB2 and phospho-Erk1/2, phospho-Akt; RT-qPCR for expression analysis; wound-healing and Transwell migration/invasion assays; immunofluorescence for focal adhesion markers (paxillin); co-immunoprecipitation of EFNB2?CEphB4 complexes; apoptosis assays; RNA-seq for transcriptomic profiling; and high-throughput genetic screens for synthetic lethal interactions or drug target validation. For further information or to discuss tailored experimental protocols, please contact Ascent Research.
