DISC1 Knockout U-251MG Cell Line

The DISC1 Knockout U-251MG Cell Line is a CRISPR/Cas9-edited cell line derived from the human U-251MG glioblastoma line, featuring targeted disruption of the DISC1 gene. This knockout model enables stable loss-of-function studies of the DISC1 scaffold protein, circumventing the limitations of transient gene silencing. The cell line is suitable for investigating DISC1-dependent signaling and cellular phenotypes, providing a robust platform for both mechanistic and translational research.

U-251MG is a widely used human glioblastoma cell line originally established from a grade IV astrocytoma. It exhibits a mesenchymal-like phenotype with high invasive and tumorigenic capacity, making it a relevant model for glioblastoma multiforme. The line retains key oncogenic signaling alterations, including dysregulation of growth factor and cytoskeletal pathways, offering a permissive background for studying the contributions of neurodevelopmental proteins to glioblastoma progression.

DISC1 encodes a multifunctional scaffold protein that assembles signaling complexes regulating neurodevelopment. It receives signals from upstream factors such as BDNF, Reelin, GSK3??, AKT, and PKA, and coordinates downstream effectors including NDE1/NDEL1, LIS1, PDE4, and ERK1/2. DISC1 directly interacts with FEZ1, ATF4, MAP1A, TNIK, and Kalirin-7, modulating pathways like Wnt/GSK3??, cAMP/PKA, and PI3K/AKT. Through these interactions, DISC1 controls processes such as neuronal migration and neurite outgrowth, and its disruption alters phosphorylation dynamics and transcriptional programs.

Loss of DISC1 in U-251MG cells disrupts the integration of neurodevelopmental signaling nodes that may intersect with glioblastoma pathology. Since DISC1 scaffolds interactions between AKT, GSK3??, and NDE1/LIS1 complexes, its knockout can perturb cytoskeletal organization, cell migration, and invasion??traits critical to glioma aggressiveness. This model therefore allows dissection of how scaffold proteins influence mesenchymal-like tumor cell behavior and may uncover signaling dependencies that are amenable to pharmacological intervention.

Researchers can employ this knockout cell line for diverse applications, including Western blot analysis of DISC1 and phospho-proteins (e.g., phospho-AKT, GSK3??), RT-qPCR for gene expression changes, and immunofluorescence to visualize interacting partners. Functional assays like Boyden chamber migration, invasion, and MTT proliferation can be conducted to assess phenotypic consequences. The model is suited for kinase inhibitor screening, functional complementation, and co-immunoprecipitation of DISC1 complexes. It also serves as a tool for psychiatric disease mechanism studies and transcriptomic analysis by RNA-seq. For additional details, please contact Ascent Research.