The DPYSL2 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human non-small cell lung carcinoma epithelial cell line, featuring targeted disruption of the DPYSL2 gene (encoding collapsin response mediator protein 2, CRMP2). As a polyclonal pool, this model provides a heterogeneous population with DPYSL2 loss-of-function, avoiding the biases of clonal selection. It is well-suited for robust investigations of DPYSL2-dependent signaling pathways in a lung adenocarcinoma background.
The A-549 host cell line is a standard model for lung adenocarcinoma, originally isolated from a male patient. These epithelial cells carry a KRAS G12S activating mutation and express wild-type p53, making them highly relevant for studying KRAS-driven oncogenic signaling in a p53-proficient context. A-549 cells are widely used in drug metabolism studies and in vitro NSCLC research, offering a reliable system to evaluate targeted therapies and the molecular mechanisms of lung cancer progression, including cytoskeletal regulation and metastatic behavior.
DPYSL2/CRMP2 is a multifunctional phosphoprotein that orchestrates cytoskeletal dynamics, cell migration, and axonal guidance. It directly interacts with ??/??-tubulin, F-actin, endophilin-A1, kinesin-1, and phosphatidylinositol (4,5)-bisphosphate to control microtubule assembly and stability, actin filament dynamics, and endocytic trafficking. Its activity is tightly regulated by upstream kinases such as GSK3??, CDK5, and ROCK, as well as by semaphorin-3A/neuropilin-1/plexin-A signaling and BDNF. Notably, GSK3??-mediated phosphorylation of CRMP2 promotes axonal growth cone collapse and modulates microtubule dynamics. Downstream, CRMP2 activates Rac1 and coordinates endophilin-A1-mediated endocytosis, thereby integrating signals to drive cytoskeletal remodeling essential for cell motility.
In A-549 lung adenocarcinoma cells, which harbor the KRAS G12S mutation driving PI3K-Akt and Rho GTPase signaling, DPYSL2 knockout disrupts microtubule stability and endocytosis, processes critically involved in cancer cell invasion and metastasis. DPYSL2 likely functions as a key effector linking oncogenic KRAS signals to cytoskeletal reorganization. Thus, this knockout model enables precise dissection of CRMP2-mediated pathways that contribute to NSCLC metastatic potential, allowing researchers to assess impacts on tumor cell migration, adhesion, and response to chemotherapeutic agents, and to identify potential therapeutic vulnerabilities within the KRAS-DPYSL2 axis.
Typical research applications include wound healing and transwell invasion assays to quantify cell migration and invasiveness, western blotting for phosphorylated DPYSL2, co-immunoprecipitation of tubulin and actin to probe protein interactions, and microtubule polymerization assays. Additionally, RNA-seq analysis and immunofluorescence for cytoskeletal structures enable comprehensive pathway profiling. These applications support drug sensitivity screening and the validation of candidate drugs targeting cytoskeletal regulators. For further information or technical support, please contact Ascent Research.
