The AP3S2 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from Jurkat T lymphocytes, with targeted disruption of the AP3S2 gene. This heterozygous pool of cells eliminates functional AP3S2 protein, enabling loss-of-function studies without clonal isolation. The genetic modification disrupts the sigma2 subunit of adaptor protein complex 3 (AP-3), a clathrin-associated complex that sorts cargo from the trans-Golgi network to lysosomes and lysosome-related organelles.
Jurkat cells are a human T-cell leukemia line (clone E6-1) established from a 14-year-old male with acute lymphoblastic leukemia. Widely used as a model for T-cell receptor (TCR) signaling and activation, these suspension cells provide a tractable system to investigate how lysosomal trafficking impacts immune cell function.
AP3S2 encodes the ??2 subunit, which assembles with AP3B1, AP3D1, and AP3M1 to form the heterotetrameric AP-3 complex. This complex interacts with clathrin and the GTPase ARF1 to mediate vesicular transport of transmembrane proteins such as LAMP1, LAMP2, and CD63. The gene is transcriptionally regulated by TFEB, a master controller of lysosomal biogenesis, and is influenced by mTORC1 signaling. AP3S2 knockout disrupts AP-3 complex integrity, causing mislocalization of lysosomal membrane proteins to the plasma membrane, impaired lysosomal acidification, and reduced degradative capacity.
In Jurkat T cells, AP-3 dysfunction mimics aspects of Hermansky-Pudlak syndrome type 2 and has been linked to neurodevelopmental disorders with epilepsy. This model allows dissection of how lysosomal trafficking defects alter TCR signaling, cytokine secretion, and immune activation. It also offers a platform to study lysosomal contributions to leukemia cell biology and to explore therapeutic interventions targeting AP-3-related pathways.
Applications include western blotting and immunofluorescence for LAMP1/LAMP2, flow cytometry for surface-exposed lysosomal proteins, lysosomal pH measurement, and co-immunoprecipitation of AP-3 subunits. The polyclonal knockout cells are suitable for genetic screens, drug discovery programs targeting lysosomal pathways, and disease modeling. For technical inquiries, contact Ascent Research.
