The ITSN2 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the ITSN2 gene in a human T-lymphocyte background. This heterogeneous pool of Jurkat cells with targeted disruption of ITSN2 enables interrogation of gene function without clonal selection biases. The polyclonal format maintains population heterogeneity, reflecting a more physiological range of knockout efficiencies, and is well-suited for pooled functional screens and bulk biochemical analyses.
The parental Jurkat cell line is an immortalized human T lymphocyte from a 14-year-old male with T-cell acute lymphoblastic leukemia (T-ALL). Jurkat cells lack functional PTEN, resulting in constitutive PI3K/AKT activation that mimics oncogenic signaling in many T-ALL cases. Widely used for T-cell receptor (TCR) signaling studies, they retain key proximal components including LCK, ZAP70, and downstream MAPK cascades.
ITSN2 encodes an endocytic adaptor protein and a guanine nucleotide exchange factor (GEF) for CDC42. Upon TCR activation, ITSN2 is recruited to the receptor complex, where it interacts with dynamin (DNM1/DNM2) and synaptojanin-1 to facilitate clathrin-mediated TCR internalization. Simultaneously, ITSN2 activates CDC42, triggering N-WASP?CArp2/3-mediated actin branching and F-actin polymerization. This dual role bridges endocytic machinery and cytoskeletal remodeling, modulating signal amplitude downstream of LCK and ZAP70. ITSN2 also interacts with SOS1, GRB2, and PI3K, connecting it to RAS?CMAPK/ERK and PI3K?CAKT pathways.
In Jurkat cells, ITSN2-mediated regulation of TCR internalization and actin dynamics is critical for immunological synapse formation and maintenance. Loss of ITSN2 is expected to alter TCR downregulation kinetics and disrupt the balance between signaling and endocytic trafficking, potentially affecting activation markers such as CD69 and CD25. With hyperactive PI3K signaling due to PTEN deficiency, the knockout model enables dissection of crosstalk between endocytic control and oncogenic pathways, providing insight into how adaptor proteins coordinate immune receptor trafficking in leukemic T cells.
This knockout model supports diverse applications including Western blotting for ITSN2, phospho-ZAP70, and phospho-ERK; flow cytometric analysis of CD69 and CD25; immunofluorescence microscopy for ITSN2 and F-actin co-localization; and transferrin uptake assays for endocytosis efficiency. Co-immunoprecipitation with dynamin-2 and real-time cell migration assays further validate disrupted interactions and functional consequences. These cells are suitable for functional screening of ITSN2-interacting partners in leukemia and drug target validation in T-ALL. For additional information or to request a quote, please contact Ascent Research.
