The DPYSL5 Knockout NCI-H1299 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DPYSL5 gene in the human non-small cell lung carcinoma cell line NCI-H1299. This loss-of-function model enables investigation of collapsin response mediator protein 5 (CRMP5) function in a cancer cell context. The polyclonal knockout pool provides a heterogeneous cell population with disrupted DPYSL5 expression, suitable for functional studies without clonal selection bias. This product serves as a versatile tool for dissecting DPYSL5-dependent signaling and cellular processes.
NCI-H1299 is a widely used human non-small cell lung carcinoma epithelial cell line derived from lymph node metastasis of a lung adenocarcinoma. It is p53-deficient and tumorigenic in nude mice, making it a robust model for studying lung cancer pathogenesis, particularly processes related to tumorigenesis, metastasis, and drug resistance. The cell line’s aggressive phenotype and well-characterized signaling networks provide a physiologically relevant platform for evaluating gene function in advanced lung cancer. This background is especially suited for examining genes implicated in cytoskeletal dynamics and cellular motility, because NCI-H1299 cells exhibit invasive characteristics in vitro and in vivo.
DPYSL5 encodes CRMP5, a member of the collapsin response mediator protein family that plays a critical role in actin cytoskeleton reorganization and microtubule assembly. CRMP5 acts downstream of semaphorin-3A (SEMA3A) signaling, where it is regulated by phosphorylation through upstream kinases such as cyclin-dependent kinase 5 (CDK5) and glycogen synthase kinase-3 beta (GSK3B). In the canonical pathway, SEMA3A binding to neuropilin-1 (NRP1) and plexin-A1 (PLXNA1) activates FYN, leading to CRMP5 phosphorylation and subsequent cytoskeletal remodeling. CRMP5 interacts with tubulin, CRMP1, CRMP2, and Ca2+/calmodulin-dependent protein kinase II (CaMKII), and modulates RhoA activity to coordinate actin dynamics. This signaling nexus regulates growth cone collapse in neurons, but in non-neuronal cells, it influences cell migration and adhesion through similar cytoskeletal mechanisms.
In the context of NCI-H1299 lung cancer cells, DPYSL5 knockout is predicted to impair semaphorin-mediated cytoskeletal reorganization, potentially affecting cell migration, invasion, and metastatic behavior. Given that NCI-H1299 cells are metastatic in origin, loss of DPYSL5 may alter their motility and invasiveness, providing insights into CRMP5??s role in cancer progression. CRMP5 has been implicated in paraneoplastic neurological syndromes, where it acts as an autoantigen; thus, this model may also help elucidate tumor-immune interactions. The combination of a tumorigenic background and DPYSL5 disruption creates a unique system to explore how neuronal guidance cues are repurposed in cancer cells to drive motility and metastasis.
This polyclonal knockout cell pool is suitable for a wide range of functional assays, including Transwell migration and invasion assays to measure metastatic potential, immunofluorescence staining to visualize actin cytoskeletal changes, and phospho-signaling analysis of CRMP5 and its interactors. Co-immunoprecipitation with tubulin or other CRMP family members can probe protein-protein interactions. Transcriptome analysis by RNA-seq can uncover global expression changes upon DPYSL5 loss, while drug sensitivity studies may reveal synthetic vulnerabilities. The model facilitates investigation of semaphorin signaling in non-neuronal cells and evaluation of DPYSL5 as a therapeutic target. For detailed technical specifications and ordering information, please contact Ascent Research.
