The OCRL Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the OCRL gene, providing a versatile loss-of-function model for investigating OCRL biology. This polyclonal pool offers a heterogeneous knockout background, enabling robust functional studies without clonal isolation, and is suitable for diverse downstream applications in cell biology and disease research.
The host Raji cell line is a human B lymphocyte derived from Burkitt lymphoma and is persistently positive for Epstein-Barr virus. These cells maintain immune effector functions, including antibody production, and are widely utilized as a model system for studying endocytosis, signal transduction, and lymphocytic malignancies. Their rapid proliferation and genetic tractability make them an ideal platform for gene editing.
OCRL encodes a phosphatidylinositol 4,5-bisphosphate 5-phosphatase that hydrolyzes PI(4,5)P2 on endosomal membranes, a critical step in coordinating membrane trafficking with actin cytoskeleton remodeling. OCRL is activated downstream of the small GTPases Rab5 and Rab35, and its activity is modulated by PI3K and PIP5K. Through direct interactions with adaptor proteins APPL1, GIPC1, clathrin, IPIP27A, IPIP27B, and the AP2 complex, OCRL regulates local PI(4,5)P2 levels to control endosomal sorting. In turn, this governs actin polymerization via Rac1, Cdc42, and the Arp2/3 complex, and is essential for primary cilia assembly. Loss of OCRL leads to PI(4,5)P2 accumulation, aberrant actin dynamics, disrupted endosomal signaling, and defective ciliogenesis, which are hallmarks of Lowe syndrome and Dent disease 2.
In the Raji B lymphocyte context, OCRL knockout disrupts endocytic trafficking and actin organization, providing a human disease-relevant model to study Lowe syndrome pathology. Because Raji cells exhibit active endocytosis and membrane recycling, this knockout population is particularly suited for dissecting the integration of phosphoinositide signaling with cytoskeletal regulation in immune cells, and for exploring the consequences of PI(4,5)P2 dysregulation on cellular functions.
This OCRL knockout polyclonal cell population is ideal for disease modeling of Lowe syndrome and ciliopathies, as well as for mechanistic studies of endocytosis and actin dynamics. Key assays include transferrin uptake for endocytic efficiency, immunofluorescence for acetylated tubulin to assess cilia formation, and PI(4,5)P2 ELISA to quantify lipid levels. Further applications encompass drug screening for PI3K-related disorders and co-immunoprecipitation to probe protein interactions. For additional information, please contact Ascent Research.
