The DPYSL2 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa human cervical adenocarcinoma cell line. This product features targeted disruption of the DPYSL2 gene, which encodes collapsin response mediator protein 2 (CRMP2), a multifunctional phosphoprotein pivotal for microtubule assembly, stabilization, and cytoskeletal remodeling. The polyclonal format comprises a heterogeneous mixture of cells carrying diverse CRISPR/Cas9-induced mutations, thereby enabling robust loss-of-function analyses while avoiding the biases associated with clonal selection. These cells provide a versatile research tool for dissecting CRMP2-dependent processes in a well-established epithelial cancer model.
HeLa cells are an adherent, HPV18-positive, aneuploid cervical carcinoma line that has been extensively characterized in cancer biology. Their robust proliferation, well-mapped signaling networks, and transformed phenotype make them highly suitable for investigating the non-neuronal functions of neuronal guidance molecules such as CRMP2. The invasive properties of HeLa cells offer a physiologically relevant system to evaluate how DPYSL2 loss impacts tumor cell migration, adhesion, and metastatic potential within the context of cervical adenocarcinoma.
DPYSL2 acts downstream of semaphorin 3A, neuropilin-1, and plexin-A receptors, integrating extracellular guidance cues through phosphorylation by kinases including GSK3??, CDK5, and Fyn. Once activated, CRMP2 binds directly to tubulin heterodimers to promote microtubule assembly and stabilization, while its phosphorylation reduces this affinity, leading to cytoskeletal collapse. CRMP2 also interacts with actin filaments, kinesin light chain 1, LIS1, and Numb, thereby coordinating microtubule dynamics with actin rearrangement and intracellular transport. Knockout of DPYSL2 disrupts these multi-protein interactions, impairing microtubule polymerization, actin network organization, and semaphorin-mediated cytoskeletal responses.
In HeLa cells, ablation of CRMP2 is expected to perturb the cytoskeletal architecture that underpins cell motility, division, and adhesion. The polyclonal nature of the knockout population ensures that a broad spectrum of genetic disruptions is represented, averaging out clonal outliers and revealing the core functions of CRMP2 in cervical carcinoma. This model system is particularly valuable for dissecting how semaphorin-driven pathways influence epithelial cancer cell behavior, including the regulation of cell shape and invasive capacity.
These cells are amenable to a wide array of functional assays, including Western blotting to confirm CRMP2 knockdown and assess its phosphorylation status at key regulatory sites. Immunofluorescence microscopy can visualize alterations in microtubule and actin filament architecture, while quantitative migration and invasion assays measure functional consequences of gene disruption. Co-immunoprecipitation experiments enable detection of changes in CRMP2??s interaction with tubulin and other binding partners, and phospho-signaling analysis can map downstream pathway perturbations. Additionally, this knockout model supports screening of small-molecule modulators aimed at CRMP2 phosphorylation or protein?Cprotein interactions. Researchers may employ these cells to advance understanding of non-neuronal CRMP2 roles in cancer cell dissemination, cytoskeletal dynamics, and response to extracellular cues. For further information or technical support, please contact Ascent Research.
