The DPYSL3 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the DPYSL3 gene, encoding collapsin response mediator protein 4 (CRMP4), has been disrupted. Unlike clonal lines, this heterogeneous pool captures a range of genetic alterations, allowing functional studies of gene loss in a cell population that more closely mirrors the variability found in tumor samples, making it ideal for robust migration and invasion assays.
The host HeLa cell line is an HPV18-positive human cervical adenocarcinoma line. The viral oncoproteins E6 and E7 drive degradation of p53 and inactivation of the retinoblastoma protein (Rb), respectively, resulting in loss of critical cell cycle checkpoints and genomic instability. This transformation underpins HeLa??s widespread use in cancer research for examining motility, invasion, and signaling pathways that contribute to metastasis.
DPYSL3/CRMP4 functions as a key phosphoprotein at the intersection of semaphorin and Rho GTPase signaling. It is regulated by upstream signals from Semaphorin 3A (Sema3A) through plexin receptor complexes, as well as by kinases GSK3?? and CDK5, and GTPase RhoA. CRMP4 directly binds and stabilizes microtubules and interacts with actin filaments, CRMP1/CRMP2 heterodimers, and focal adhesion kinase (FAK), integrating guidance cues to control axon growth cone collapse and cell polarization. Phosphorylation by GSK3?? and CDK5 reduces CRMP4?Ctubulin affinity, promoting microtubule disassembly and actin remodeling necessary for directional migration. Additionally, CRMP4 modulates expression of matrix metalloproteinases, connecting cytoskeletal dynamics to extracellular matrix degradation during invasion.
In HeLa cervical carcinoma cells, DPYSL3 knockout disrupts the coordinated cytoskeletal remodeling required for invasive migration, directly impairing the metastatic potential conferred by HPV-mediated transformation. Because CRMP4 expression has been correlated with migratory and invasive capacity in multiple cancer types, this knockout model provides a valuable platform to dissect the convergence of semaphorin?CCRMP signaling with HPV-altered pathways (p53/Rb) in driving tumor cell dissemination. It also allows examination of how DPYSL3 loss alters responsiveness to guidance cues like Sema3A within the tumor microenvironment.
This polyclonal knockout cell population is well-suited for a variety of experimental approaches: transwell migration and wound healing assays to measure directional motility, immunofluorescence visualization of F-actin and tubulin networks to assess cytoskeletal integrity, western blot analysis of total and phosphorylated CRMP4 levels, co-immunoprecipitation to map protein-protein interactions with FAK or tubulin, and RNA-seq transcriptomic profiling to uncover downstream gene expression changes. The cells facilitate detailed investigation of semaphorin?Cplexin signal transduction, RhoA?CROCK-mediated actin reorganization, and microtubule dynamics in cancer cell migration and invasion. For further inquiries or technical assistance, please contact Ascent Research.
