The DPYSL5 Knockout 786-O Polyclonal Cells consist of a polyclonal population of 786-O cells subjected to CRISPR/Cas9-mediated disruption of the DPYSL5 gene, encoding collapsin response mediator protein 5 (CRMP5). As a polyclonal pool, this product delivers a heterogeneous knockout model that reflects the natural variability of genetic edits, enabling robust loss-of-function studies without clonal artifacts. The cells are provided as a ready-to-use frozen stock for immediate culture and downstream applications.
The host 786-O cell line, isolated from a primary clear cell renal adenocarcinoma of a 58-year-old female, harbors a well-documented VHL F148L mutation, which abrogates HIF-?? degradation and establishes a constitutive pseudo-hypoxic state. This drives an angiogenic and highly invasive phenotype, positioning 786-O as a cornerstone model in renal cell carcinoma (RCC) research, particularly for investigating the interplay between hypoxia signaling and tumor progression.
DPYSL5 (CRMP5) is a phosphoprotein that operates at the intersection of semaphorin and reelin signaling pathways. Upon stimulation by SEMA3A through NRP1/PLXNA1 receptors or activation by reelin, CRMP5 is phosphorylated by upstream kinases including GSK3??, CDK5, ROCK, and FYN, which dictates its ability to bind microtubule and actin cytoskeletal components. Through direct interactions with TUBB, ACTB, and VIM, CRMP5 orchestrates microtubule polymerization and actin filament reorganization, while also modulating the activity of small GTPases RAC1 and RHOA. This molecular rheostat function is critical for processes like growth cone collapse in neurons and for the directed migration and invasion of tumor cells.
In the context of 786-O cells with constitutive HIF activation, loss of DPYSL5 disrupts the cytoskeletal reorganization required for tumor cell motility and invasiveness. By impairing CRMP5-mediated coordination of microtubule dynamics and actin remodeling, the knockout model attenuates the invasive machinery downstream of pro-migratory semaphorin cues. This makes it a valuable system for studying the cytoskeletal basis of RCC metastasis and for exploring the role of CRMP5 in paraneoplastic neurological disorders, where it is a common autoantigen.
This DPYSL5 polyclonal knockout product is designed for use in a range of functional assays, including Transwell migration and invasion, wound healing, and Boyden chamber experiments to assess motility and matrix degradation. Complementary molecular analyses such as immunofluorescence staining for tubulin and actin, Western blotting for total and phospho-CRP5, and co-immunoprecipitation of tubulin complexes allow detailed examination of CRMP5-dependent cytoskeletal changes. Live-cell imaging of cytoskeletal dynamics and Rho GTPase activation assays further enable real-time pathway dissection. Key applications include renal cell carcinoma functional genomics, semaphorin-plexin signaling studies, cytoskeletal drug target validation, and epithelial-mesenchymal transition research. For additional information or custom requests, please contact Ascent Research.
