The Nucks1 Knockout HT22 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the mouse hippocampal neuronal HT22 cell line, in which the Nucks1 gene has been disrupted. This loss-of-function model enables investigation of Nucks1-dependent cellular processes, including DNA repair, cell cycle regulation, and transcriptional control. The knockout cell line provides a defined genetic background for studying the role of Nucks1 in neuronal stress responses and neurodegenerative pathways.
HT22 is an immortalized mouse hippocampal neuronal cell line, originally a subclone of HT4, widely employed as a model for neuronal oxidative stress and glutamate-induced toxicity. HT22 cells lack functional ionotropic glutamate receptors, undergoing oxidative glutamate toxicity via cystine/glutamate antiporter inhibition, which mimics excitotoxicity-independent neuronal death. This makes HT22 particularly valuable for studying neurodegeneration mechanisms mediated by oxidative stress, a key feature in Parkinson??s disease and other disorders.
Nucks1 (Nuclear Casein Kinase and Cyclin-Dependent Kinase Substrate 1) is a nuclear phosphoprotein that functions at the intersection of DNA damage response and transcriptional regulation. It is phosphorylated by CK2 and CDKs (CDK1, CDK2) upon DNA damage, acting downstream of ATM/ATR signaling. Nucks1 transcriptionally regulates the alpha-synuclein gene SNCA, a central player in Parkinson??s disease pathology, as well as cell cycle and apoptosis genes such as CDKN1A (p21) and BAX. Through these interactions, Nucks1 modulates cell cycle checkpoint control and apoptosis in response to genotoxic stress.
In the HT22 neuronal background, Nucks1 knockout significantly alters cellular responses to oxidative and genotoxic insults. The loss of Nucks1 impairs DNA damage repair and cell cycle checkpoint activation, while reducing SNCA expression, which may influence alpha-synuclein aggregation dynamics. This model thus recapitulates molecular alterations observed in Parkinson??s disease and other neurodegenerative conditions, offering a platform to dissect how Nucks1-dependent pathways affect neuronal vulnerability to stress.
Applications include neurodegenerative disease research, DNA damage response studies, and apoptosis assays. Researchers can employ Western blotting, RT-qPCR, and immunofluorescence to quantify changes in SNCA, p21, BAX, and phospho-ATM/ATR. Flow cytometry enables cell cycle profiling and apoptosis detection, while the comet assay measures DNA damage accumulation. Glutamate-induced oxidative stress assays using this knockout line reveal the role of Nucks1 in neuronal survival. For further information or technical support, please contact Ascent Research.
