The Ulk4 Knockout Neuro-2a Cell Line is a CRISPR/Cas9-edited mouse neuroblastoma cell line designed to disrupt the Ulk4 gene, enabling loss-of-function studies in a neuronal model system. This stable knockout cell line provides a valuable tool for investigating ULK4 function in autophagy and neuronal signaling pathways.
The Neuro-2a host cell line originates from a spontaneous neuroblastoma in the A/J mouse strain, displaying hallmarks of a neural crest-derived tumor. These adherent cells are commonly employed as a neuronal model due to their capacity to differentiate into neuron-like cells upon serum withdrawal or treatment with differentiation agents, expressing neuronal markers and extending neurites. This background makes them particularly relevant for studying neuronal differentiation, synaptic function, and neurodegenerative processes.
ULK4 is a serine/threonine kinase with key roles in autophagy initiation and neuronal development. It is posited to act downstream of the nutrient sensors mTORC1 and AMPK, which phosphorylate ULK4 to modulate its activity in response to cellular energy status. As part of the autophagy initiation complex, ULK4 interacts with ATG13, FIP200, and the ULK1/2 kinases, potentially scaffolding the assembly of the phagophore. Downstream effects include regulation of the Beclin-1?CVPS34 lipid kinase complex and LC3 lipidation, ultimately influencing autophagosome formation. In neurons, ULK4 also impacts neurite outgrowth and synaptic homeostasis, linking autophagy to neuronal architecture.
Disruption of Ulk4 in Neuro-2a cells offers insight into the intersection of autophagy and neuronal biology. Since these cells undergo neuronal differentiation, the knockout model permits examination of ULK4??s role in neurite extension and stress responses. Given genetic associations with schizophrenia, bipolar disorder, and autism spectrum disorder, this line aids research into the molecular mechanisms of neuropsychiatric disorders and autophagy-related neuronal dysfunction.
Researchers can utilize this knockout cell line for detailed functional studies. Representative assays include Western blotting for LC3 and p62 to monitor autophagy flux, immunofluorescence microscopy to visualize autophagosome markers like LC3 puncta, RT-qPCR to quantify expression of neuronal genes, viability assays under metabolic or proteotoxic stress, neurite outgrowth measurements, mTOR activity determination via phospho-S6 immunoblotting, flow cytometry with fluorescent reporters for autophagy flux, co-immunoprecipitation to identify ULK4-containing protein complexes, and drug sensitivity screening for compounds that modulate autophagy or neuronal phenotypes. For further information or to order, please contact Ascent Research.
