The EFNA5 Knockout Jurkat Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout population derived from Jurkat T lymphocytes, designed to disrupt the EFNA5 gene. This pool provides a loss-of-function model for ephrin-A5, a GPI-anchored EphA receptor ligand, enabling studies of its role in T-cell signaling without clonal selection artifacts. The heterogeneous edited alleles within the polyclonal pool enable robust functional analysis of ephrin-A5-dependent processes in T-cell biology and oncogenic signaling.
Jurkat cells are an immortalized human T-lymphocyte line derived from an acute T-cell leukemia patient. Widely used for studying TCR signaling, apoptosis, and leukemia biology, these suspension cells proliferate rapidly and are easily manipulated genetically. Their leukemic background is particularly relevant for investigating aberrant signaling pathways driving malignant transformation. The EFNA5 knockout Jurkat pool thus offers a physiologically pertinent model to examine how ephrin-A5 loss affects T-cell adhesion, migration, and receptor-mediated communication.
EFNA5 encodes ephrin-A5, a membrane-tethered ligand that binds EphA receptors (EphA3, EphA4, EphA8) to initiate bidirectional signaling. EphA forward signaling activates Src and Abl kinases, modulating RhoA/ROCK and MAPK/ERK pathways that control cytoskeletal dynamics and adhesion. Ephrin-A5 is cleaved by ADAM10 to terminate adhesion. Expression is regulated by retinoic acid, Notch, and Wnt. The ephrin-A5/EphA system mediates contact-dependent repulsion and adhesion critical for axon guidance and tissue morphogenesis, with dysregulation linked to cancer metastasis and glioblastoma.
In Jurkat T cells, ephrin-A5 intersects with integrin-mediated adhesion and migration pathways, likely influencing leukemic cell behavior. EFNA5 disruption abrogates EphA receptor activation, uncoupling Rho GTPase and ERK1/2 signaling. This model allows dissection of ephrin-A5’s role in T-cell repellent responses and assessment of altered phosphorylation of effectors like ERK1/2 and RhoA. With relevance to immune cell trafficking and tumor dissemination, these cells facilitate studies of leukemia cell migration and adhesion plasticity.
Typical applications include transwell migration and cell adhesion assays using EphA-Fc substrates, flow cytometry for EphA receptor expression, and western blotting for phospho-ERK1/2 or RhoA activity. Co-immunoprecipitation confirms disrupted EphA-ephrin complexes. These cells also support synaptic plasticity and developmental neurobiology research due to ephrin-A5’s neuronal roles. For further details, please contact Ascent Research.
