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Voltage-gated sodium channel α-subunit type Ⅲ (Nav1.3) is mainly expressed in the central nervous system and is associated with neurological disorders.The expression of mouse Scn3a product (Nav1.3) mainly occurs in embryonic and early postnatal brain but not in adult brain.Here, we report for the first time the identification and characterization of the mouse Scn3a gene promoter region and regulation of the promoter activity by GC box and CpG methylation.Luciferase assay showed that the promoter region F1.2 (nt-1,049 to +157) had significantly higher activity in PC 12 cells, omparing with that in SH-SY5Y cells and HEK293 cells.A stepwise 5 truncation of the promoter region found that the minimal functional promoter located within the region nt-168 to +157.eletion of a GC box (nt-254 to-258) in the mouse Scn3a promoter decreased the promoter activity.CpG methylation of the F1.2 without the GC box completely repressed the promoter activity, suggesting that the GC box is a critical element in the CpG-methylated Scn3a promoter.These results suggest that the GC box and CpG methylation might play important roles in regulating mouse Scn3a gene expression.