The DPYSL5 Knockout NCI-H1975 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the DPYSL5 gene in the NCI-H1975 human lung adenocarcinoma epithelial cell line. This polyclonal knockout format provides a heterogeneous pool of gene-edited cells, eliminating the need for single-cell cloning and enabling robust loss-of-function analysis. The targeted disruption of DPYSL5, which encodes collapsin response mediator protein 5 (CRMP5), abrogates expression of this critical cytoskeletal regulator, allowing researchers to dissect its roles in cancer cell biology.
The NCI-H1975 host cell line was originally derived from the lung adenocarcinoma of a non-smoking female and is characterized by well-defined oncogenic mutations, including EGFR exon 21 L858R and T790M point mutations, as well as a PIK3CA G118D substitution. This genetic profile renders the cells sensitive to first- and second-generation EGFR tyrosine kinase inhibitors while modeling acquired resistance, making them a highly relevant in vitro system for non-small cell lung cancer (NSCLC) research, particularly in the context of targeted therapeutic approaches.
CRMP5 functions as a pivotal mediator of semaphorin-3A (SEMA3A) signaling, acting downstream of the neuropilin-1/plexin-A receptor complex. Upon ligand engagement, GSK3??-mediated phosphorylation of CRMP5 modulates its interactions with tubulin heterodimers and the dynein motor complex, thereby orchestrating microtubule polymerization and actin cytoskeleton reorganization. This signaling cascade converges on small GTPases RhoA and Rac1, which govern directed cell migration and invasion. Consequently, DPYSL5 knockout disrupts SEMA3A-induced cytoskeletal remodeling, providing a defined loss-of-function model to interrogate this pathway.
Within the NCI-H1975 background harboring concurrent EGFR and PIK3CA pathway activation, DPYSL5 depletion is expected to unmask the contribution of CRMP5-dependent cytoskeletal dynamics to oncogene-driven phenotypes. By uncoupling semaphorin-mediated cytoskeletal control from parallel MAPK and PI3K signaling, this model enables investigation of how microtubule reorganization influences lung adenocarcinoma progression, epithelial-mesenchymal transition (EMT), and the emergence of drug-resistant cell states. Such studies are critical for understanding metastasis in EGFR-mutant NSCLC.
This polyclonal knockout product is well-suited for functional assays that quantify cell migration and invasion, including scratch wound healing and transwell assays, complemented by immunofluorescence imaging of the microtubule and actin networks. Western blot analysis of phosphorylated CRMP5 and co-immunoprecipitation experiments can confirm disrupted protein complexes, while RT-qPCR profiling of EMT markers (e.g., CDH1, VIM) connects cytoskeletal changes to transcriptional programs. Additional applications encompass flow cytometric cell cycle analysis, microtubule polymerization kinetics assays, and drug sensitivity profiling against EGFR inhibitors. For detailed technical data and ordering information, please reach out to Ascent Research.
