The DOCK2 Knockout UM-UC-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the UM-UC-3 human bladder carcinoma line. This loss-of-function model, generated by targeted disruption of the DOCK2 gene, provides a heterogeneous pool of alleles for robust functional studies without clonal selection.
The UM-UC-3 cell line is derived from a transitional cell carcinoma of the human bladder and is extensively used as a model for invasive urothelial carcinoma. These epithelial cells harbor a mutation in the TP53 tumor suppressor gene and exhibit a mesenchymal phenotype associated with enhanced migratory and invasive capabilities. UM-UC-3 cells are adherent, proliferate robustly in standard culture media, and are amenable to transfection, lentiviral transduction, and CRISPR-based genome editing, making them a reliable platform for knockout studies.
DOCK2 is a Rac-specific guanine nucleotide exchange factor predominantly expressed in hematopoietic cells, where it is essential for chemokine-induced migration and lymphocyte activation. DOCK2 catalyzes the conversion of inactive GDP-bound RAC1 and RAC2 to their active GTP-bound forms. Upon activation, RAC stimulates actin polymerization and lamellipodia formation through downstream effectors including PAK1, the WAVE2 complex, and the ARP2/3 complex. DOCK2 acts downstream of chemokine receptors (e.g., CXCR4, CCR7) and is regulated by the scaffold proteins ELMO1 and DOCK2BP, which facilitate its membrane recruitment and catalytic activity. The canonical CXCL12-CXCR4-DOCK2-ELMO1-RAC1 signaling pathway drives directed cell migration, while DOCK2 also participates in T cell receptor and B cell receptor signaling for immune synapse formation.
While DOCK2 is primarily studied in immune cells, its knockout in the UM-UC-3 background enables investigation of potential non-canonical roles in bladder cancer biology. The p53-null, mesenchymal context provides a model to explore whether DOCK2 influences epithelial cell motility, invasion, or tumor-immune microenvironment interactions. These cells serve as a tool for co-culture studies to assess the contribution of tumor-intrinsic DOCK2 to immune cell recruitment and activation.
Researchers can employ these polyclonal DOCK2 knockout cells in a variety of functional assays. Transwell migration and invasion assays are ideal for assessing chemotactic responses, while Rac-GTP pull-down experiments provide direct readouts of DOCK2-dependent RAC activation. F-actin staining with phalloidin and immunofluorescence microscopy can visualize cytoskeletal rearrangements. The cells are suitable for flow cytometric analysis of surface markers, co-culture with immune cells to study tumor-immune interactions, and Western blotting for signaling molecules such as phospho-PAK1. The polyclonal format also facilitates pooled CRISPR screens and synthetic lethality studies. For technical support and ordering, please contact Ascent Research.
