mOKT3-scFv[Puro-GFP] Lentivirus

The mOKT3-scFv[Puro-GFP] lentivirus is a research-grade, integratable vector designed to deliver a membrane-bound single-chain variable fragment (scFv) derived from the murine OKT3 monoclonal antibody, which targets CD3, and contains a bicistronic selection and reporter gene cassette (puromycin-2A-GFP). This system leverages the established advantages of lentiviral vector delivery, enabling efficient gene transfer into a wide range of mammalian cells, including difficult-to-transfect primary cells, and achieving stable expression through genomic integration. The membrane-bound OKT3 scFv is engineered for stable expression on the cell surface, allowing for precise localized binding to CD3 on T cells while minimizing dilution effects caused by secretion. The inclusion of a self-cleaving 2A peptide ensures coordinated co-expression of puromycin resistance and GFP from the same transcript, simplifying antibiotic selection of stable cell populations/clones and allowing for real-time visualization or enrichment of transduced cells via fluorescence.

This vector is ideally suited for immunology and cell engineering studies requiring controlled T cell activation via CD3. Typical applications include the construction of artificial antigen-presenting cells (aAPCs) or feeder cell lines that provide sustained CD3 stimulation in co-culture models for T cell activation, expansion, or research. It can support mechanistic studies of T cell receptor signaling, immunological synapse formation, and downstream transcriptional or functional responses under specific conditions. Engineered tumor cells or stromal cells expressing membrane-bound OKT3 scFv can serve as a customizable platform for exploring T cell-tumor interactions, screening for modulators of T cell activation, or evaluating signaling in combination with transgenically introduced co-stimulatory ligands. The GFP reporter gene facilitates rapid assessment of transduction efficiency, live-cell imaging of engineered cell populations, and fluorescence-based enrichment when used in conjunction with flow cytometry, while puromycin resistance aids in establishing stable cell lines for long-term studies.