The CCDC91 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat T lymphoblast cell line, designed for targeted disruption of the CCDC91 gene. This product offers a heterogeneous pool of edited cells, generated through electroporation of CRISPR/Cas9 ribonucleoproteins, providing a versatile loss-of-function model for studying the molecular functions of the coiled-coil domain-containing protein 91. Unlike clonal lines, the polyclonal nature maintains genetic diversity, enabling robust population-level analyses of CCDC91-dependent phenotypes without clonal selection artifacts. Researchers can employ this model to systematically dissect retrograde trafficking from endosomes to the trans-Golgi network and evaluate pathways regulating lysosomal homeostasis in T cell leukemia contexts.
Jurkat cells are an extensively characterized human T lymphoblast cell line originally isolated from the peripheral blood of a 14-year-old male with relapsed acute T cell leukemia. These immortalized cells constitutively express the T cell receptor (TCR)?CCD3 complex and proliferate in an interleukin-2-independent manner, making them a standard model for studying TCR signaling, activation dynamics, and HIV infection. Their leukemic origin and fully functional membrane trafficking machinery render them particularly suitable for investigating how vesicular sorting impacts immune receptor turnover and oncogenic signaling. The Jurkat background thus provides a pathophysiologically relevant setting for examining CCDC91??s role in clathrin-mediated endocytosis and Golgi-to-lysosome transport within a malignant T lymphocyte context.
CCDC91 encodes a coiled-coil adaptor protein that serves as a critical scaffold linking the Golgi-associated, gamma-adaptin ear-containing, ARF-binding protein 1 (GGA1) to the clathrin adaptor complex AP-1 at the trans-Golgi network (TGN). This bridging function facilitates the formation of clathrin-coated vesicles destined for endosomes and lysosomes, thereby directing the sorting of lysosomal hydrolases such as cathepsins and lysosomal membrane proteins including LAMP1 and LAMP2. CCDC91 operates downstream of the small GTPase ARF1 and upstream of cargo receptors like the cation-independent mannose-6-phosphate receptor, integrating signals from the TFEB/MITF family of transcription factors that globally control lysosomal biogenesis. Disruption of CCDC91 by CRISPR/Cas9 consequently impairs the trafficking of these degradative components, leading to defective lysosomal acidification and reduced proteolytic capacity.
In Jurkat cells, CCDC91 knockout provides a powerful tool to interrogate the intersection of membrane trafficking and T cell receptor fate. Following TCR engagement, activated receptors are internalized and sorted either for recycling back to the plasma membrane or for lysosomal degradation ?C a decision critically influenced by the endomembrane sorting machinery. Loss of CCDC91 disrupts the TGN exit of lysosomal enzymes, attenuating degradative flux and potentially altering the kinetics of TCR downregulation. This perturbation may modulate downstream signaling cascades that depend on proper receptor internalization and degradation, including those involving ZAP70, LAT, and ERK. Consequently, the knockout model helps investigate how leukemic T cells might subvert trafficking pathways to sustain surface receptor expression and proliferative signaling, illuminating potential vulnerabilities in acute T cell leukemia.
This polyclonal knockout product is suited for a spectrum of research applications, including functional genomics screens to identify trafficking regulators in leukemia, detailed mechanistic studies of clathrin adaptor function, and drug discovery efforts targeting lysosomal storage disorders or T cell malignancies. Typical assays include western blotting for CCDC91 and LAMP1, RT-qPCR to monitor transcriptional responses, immunofluorescence microscopy for colocalization of TGN and endolysosomal markers, flow cytometry to measure surface TCR levels, cathepsin enzymatic activity assays, and T cell receptor internalization kinetics. Additional techniques such as co-immunoprecipitation with GGA1 and RNA-seq transcriptome profiling can further dissect CCDC91-dependent networks. For further details, please contact Ascent Research.
