The EFNA5 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population tailored for functional interrogation of ephrin-A5. Produced by CRISPR/Cas9-mediated gene disruption in HAP1 cells, this model comprises a heterogeneous pool of EFNA5-deficient cells, avoiding the bottlenecks of clonal selection. It provides a robust population-based tool for analyzing collective cellular responses in adhesion, migration, and signaling pathways.
The parental HAP1 cell line is a near-haploid human chronic myeloid leukemia (CML) line with adherent fibroblast-like morphology, derived from the KBM-7 CML line. HAP1 cells carry the BCR-ABL fusion oncogene and are widely adopted for knockout studies owing to their haploid karyotype, which streamlines targeted gene disruption. Their hematopoietic origin and stable in vitro characteristics render them a versatile platform for generating knockout models relevant to cancer biology, neurobiology, and cell signaling research.
EFNA5 encodes ephrin-A5, a glycosylphosphatidylinositol (GPI)-anchored ligand that binds EphA receptor tyrosine kinases, including EphA3, EphA4, EphA5, and EphA7, at cell?Ccell interfaces. This engagement initiates bidirectional signaling: forward signaling through EphA receptors activates SRC, FAK, and the MAPK cascade, while reverse signaling via ephrin-A5 regulates small GTPases such as RHOA, RAC1, and CDC42 to remodel the cytoskeleton. EFNA5 transcription is modulated by upstream regulators PAX6, FGF, Wnt, and Notch pathways, and its surface availability is controlled by ADAM10-mediated proteolytic shedding. Collectively, this network orchestrates cell repulsion, axon guidance, tissue boundary establishment, angiogenesis, and synaptic plasticity.
In HAP1 cells, EFNA5 knockout eliminates ephrin-A5-dependent bidirectional communication, providing a clean genetic background to examine Eph receptor?Cmediated processes. The near-haploid nature of HAP1 ensures that a single targeted allele results in a functional null, facilitating strong phenotypic contrasts in polyclonal populations. This model is particularly suited for dissecting ephrin-A5??s contributions to adhesion-dependent signaling, cytoskeletal dynamics, migration, and proliferation, and for comparing isogenic wild-type and knockout cultures in a leukemia-derived cellular environment.
Research applications include investigating ephrin-A5??s role in neurodevelopmental disorders, axon guidance defects, and cancer progression, with emphasis on prostate and breast cancer cell migration and invasion. The polyclonal knockout pool is compatible with high-throughput genetic screens, drug target validation, and detailed biochemical analyses. Representative assays include Western blotting to confirm protein loss, co-immunoprecipitation to assess EphA receptor interactions, immunofluorescence to examine cytoskeletal reorganization, and functional assays such as transwell migration, wound-healing, and cell adhesion. For additional technical details and ordering information, please contact Ascent Research.
