Stk36 Knockout Neuro-2a Cell Line

The Stk36 Knockout Neuro-2a Cell Line is a CRISPR/Cas9-edited knockout cell line featuring targeted disruption of the mouse Stk36 gene. This product provides a defined loss-of-function model for studying serine/threonine kinase 36 (STK36) in Hedgehog signaling, utilizing the Neuro-2a neural crest-derived neuroblastoma background. The knockout line facilitates precise investigation of STK36-dependent phosphorylation events and transcriptional regulation without interference from endogenous STK36 activity.

Neuro-2a cells originate from a spontaneous tumor of an A/J mouse and serve as a widely used model for neuronal differentiation and neurobiology. These cells can be induced to differentiate into neuron-like phenotypes, making them suitable for examining Hedgehog pathway roles in neurodevelopment. The Stk36 knockout in this background enables examination of kinase function in a neuroblastoma environment relevant to Hedgehog-related neurobiology and oncogenesis.

STK36 is a serine/threonine kinase that positively regulates Hedgehog signaling. Upon pathway activation by Shh ligands and Smoothened, STK36 phosphorylates Gli1, Gli2, and Gli3 transcription factors, relieving Sufu-mediated inhibition. This phosphorylation promotes Gli transcriptional activation of targets like Gli1 and Ptch1. STK36 also interacts with kinesin proteins Kif27 and Kif7, which assist Gli protein trafficking in primary cilia. Disruption of Stk36 abolishes this critical signaling step, providing a clean background for pathway analysis.

In Neuro-2a cells, loss of Stk36 impairs Hedgehog-mediated transcriptional responses, affecting neuronal differentiation and ciliogenesis. The knockout line permits comparative studies of parental and Stk36-disrupted cells under differentiation conditions to dissect kinase contributions to neurite outgrowth and maturation. It also allows investigation of pathway cross-talk in the absence of STK36.

Research applications include screening for Hedgehog pathway modulators, western blotting for Gli phosphorylation, RT-qPCR for target genes, immunofluorescence for cilia, luciferase reporter assays, and proliferation/differentiation studies. These uses are relevant to investigations of Hedgehog-related cancers (medulloblastoma, basal cell carcinoma) and ciliopathies. For further information, researchers may contact Ascent Research.