Tub Knockout Neuro-2a Cell Line

The Tub Knockout Neuro-2a Cell Line is a CRISPR/Cas9-edited knockout cell line derived from mouse neuroblastoma Neuro-2a cells. This loss-of-function model targets the Tub gene, which encodes Tubby, facilitating studies of Tubby’s roles in neuronal signaling, metabolism, and ciliary function. Supplied as a stable knockout line, it ensures reproducible results and straightforward integration into neurobiology and metabolic disease research workflows.

Neuro-2a is a mouse neuroblastoma line originating from a spontaneous tumor of the A/J strain. It exhibits neuronal progenitor-like properties and can be differentiated into neuron-like cells with retinoic acid or serum withdrawal. The line expresses authentic neuronal markers and signaling pathways, making it a standard model for neurobiology and differentiation studies. The Tub knockout derivative retains these characteristics while ablating Tubby function, enabling gene-specific analyses in a neural context.

Tubby functions as a transcription factor and signaling adapter anchored to the plasma membrane via PtdIns(4,5)P2. Activation of Gq-coupled receptors, such as muscarinic M1, or the insulin receptor triggers PLC??-mediated hydrolysis of PtdIns(4,5)P2, releasing Tubby for importin-dependent nuclear translocation. In the nucleus, Tubby regulates transcription of metabolic genes (Ucp1, LepR), neurotransmitter receptors, and ciliary targets. It also interacts with IRS proteins and the IFT complex, integrating insulin, GPCR, and ciliary signaling.

The Neuro-2a context is ideal for probing Tubby??s neuronal functions because these cells possess endogenous insulin and GPCR signaling machinery, including PLC?? and insulin receptor. Tubby loss can impair neurite outgrowth, insulin-stimulated glucose uptake, and ciliary processes, thereby modeling aspects of retinal degeneration, obesity, and sensorineural hearing loss. This knockout line therefore links molecular signaling to neuronal pathophysiology in metabolic syndrome and ciliopathies.

Applications include mechanistic studies of Tubby-mediated transcriptional control, metabolic disease modeling, and ciliopathy investigations. Typical assays are western blotting and RT-qPCR for target validation, immunofluorescence and reporter assays for localization and activity, insulin-induced phospho-flow for signaling quantification, and neurite outgrowth or glucose uptake assays for functional readouts. The line is also suitable for drug screening in obesity and neurodegeneration research. For more information, please contact Ascent Research.