The DPYSL5 Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited heterogeneous population of HeLa cells carrying targeted disruption of the DPYSL5 locus. This polyclonal knockout model provides a loss-of-function system for studying DPYSL5 biology in an immortalized human epithelial background. The use of a polyclonal population allows assessment of bulk gene-editing effects without clonal artifacts, making it suitable for diverse functional analyses where clonal homogeneity is not required.
HeLa cells are a widely used HPV-18 positive cervical adenocarcinoma line characterized by p53 and Rb inactivation and a highly aneuploid karyotype. Their robust growth and adaptability make them a standard host for generating gene-edited knockout models. In the context of DPYSL5 disruption, HeLa cells offer a well-characterized platform to examine the gene??s impact on cytoskeletal organization, adhesion, and motility, given their epithelial origin and transformed phenotype. The altered cell cycle regulation in HeLa cells may also intersect with CRMP-dependent signaling, making them relevant for studying cancer-related functions.
DPYSL5 belongs to the collapsin response mediator protein (CRMP) family and functions downstream of the Semaphorin 3A/Neuropilin-1/Plexin-A receptor complex. Upon receptor activation, Fyn, CDK5, and GSK-3?? sequentially phosphorylate DPYSL5, leading to its detachment from microtubules and subsequent cytoskeletal collapse. This phosphorylation cascade is essential for growth cone repulsion during axon guidance. DPYSL5 also interfaces with actin dynamics by interacting with Rho GTPases including RhoA, Rac1, and Cdc42, as well as binding directly to tubulin and actin filaments. Through these interactions, DPYSL5 coordinates cytoskeletal remodeling essential for cell polarity, migration, and neuronal network formation. The phosphorylation status of DPYSL5 serves as a molecular switch that integrates upstream Semaphorin cues to control cytoskeletal dynamics.
In the HeLa cancer model, DPYSL5 knockout is particularly valuable for dissecting its contributions to tumor cell migration and invasion. The loss of DPYSL5 may alter Rho GTPase signaling and microtubule stability, thereby impacting processes such as scratch-wound closure and Transwell motility. Furthermore, the interaction of DPYSL5 with CRMP1 and CRMP2 suggests that its knockout may destabilize the CRMP complex, offering a means to study compensatory mechanisms among family members. Because HeLa cells exhibit deregulated cell cycle control and enhanced invasive potential, this knockout system enables investigation of CRMP-dependent pathways in a cancer-relevant context, potentially revealing vulnerabilities associated with neurodevelopmental and neoplastic disorders.
This product supports a range of experimental applications, including confirmation of DPYSL5 ablation by Western blotting, visualization of microtubule and actin networks via immunofluorescence, and quantitative migration/invasion assays. It is also suitable for phospho-CRMP analysis, qPCR-based transcriptomics, and high-throughput screening for modulators of Semaphorin/CRMP signaling. Additionally, these polyclonal knockout cells can be employed in neurite outgrowth assays when co-cultured with neuronal cells or used in heterologous systems to study axon guidance defects. Researchers focusing on cytoskeletal dynamics, cancer metastasis, or neurodevelopmental pathways can employ these knockout cells to interrogate DPYSL5 function in vitro. For additional information or custom orders, please contact Ascent Research.
