The DPYSL2 Knockout MES-OV Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the DPYSL2 gene in the MES-OV murine oviductal epithelial cell line. This polyclonal knockout product provides a versatile loss-of-function model for investigating the roles of its protein product, collapsin response mediator protein 2 (CRMP2), in cytoskeletal dynamics and signal transduction. The heterogeneous editing patterns within the polyclonal population mirror physiological variation, allowing robust assessment of DPYSL2-dependent phenotypes without clonal bias, in a biologically relevant oviductal epithelial context.
MES-OV cells are spontaneously immortalized oviductal epithelial cells established from Mus musculus tissue. They retain key attributes of the oviductal epithelium, including secretory activity, ciliary motility, and roles in gamete transport and early embryonic support. As an in vitro model of the oviductal microenvironment, MES-OV cells are well suited for studies of reproductive biology, inflammation, and tumorigenesis. The DPYSL2 knockout in this background thus allows for exploration of CRMP2 function in a system where epithelial integrity and secretory functions are crucial, linking semaphorin signaling to physiological processes in the female reproductive tract.
DPYSL2 encodes CRMP2, a cytosolic phosphoprotein that orchestrates microtubule polymerization and actin filament dynamics. CRMP2 functions downstream of Semaphorin-3A signaling through the Neuropilin-1/Plexin-A receptor complex. Its activity is regulated by phosphorylation by GSK-3?? and CDK5, which modulate interactions with tubulin and actin. Dephosphorylated CRMP2 promotes microtubule assembly, while phosphorylation facilitates actin reorganization via effectors such as cofilin and LIMK1. This cascade controls growth cone collapse and cell migration. In the knockout cells, disruption of DPYSL2 abrogates CRMP2 expression, impairing Sema3A-induced cytoskeletal remodeling.
In oviductal epithelium, CRMP2 maintains architecture and ciliary function through cytoskeletal regulation, and its knockout helps dissect semaphorin effects on gamete transport and embryo support. Since CRMP2 overexpression is linked to endometriosis and ovarian cancer metastasis, this model enables study of loss-of-function effects on invasiveness and migration, with the polyclonal population reflecting physiological variability.
These polyclonal knockout cells enable scratch-wound and Transwell migration/invasion assays, immunofluorescence for tubulin/F-actin, Western blotting for phospho-CRMP2, co-immunoprecipitation with tubulin, and live-cell imaging. Applications include semaphorin signaling in reproductive tissues, CRMP2 as a target in ovarian/breast cancer metastasis, neuronal differentiation, and endometriosis. Contact Ascent Research for details.
