Fig. 1

Lentivirus expressing Cdk5-sgRNA reduces CDK5 protein levels in the DS. Immunoassays were performed 14 days after LV/Cas9-sgRNA microinjection. A Schematic of the LV/Cas9-sgRNA expression vector. B HEK293 cells transduced with 10 μL LV/Cas9-Cdk5-sgRNA (2 × 10⁸ pfu/mL) and eGFP expression of HEK293-T evaluated using a fluorescence microscope. Scale bars = 200 μm. C The transfection rate of the virus, indicating a well-established effect relationship between the transfect rate and LV/Cas9-Cdk5-sgRNA virus. D Experimental timeline and schematic of microinjection sites in the mouse brain. LV/Cas9-sgRNA were highly expressed in the DS region, as indicated by GFP (green) under fluorescence microscopy. E Representative immunofluorescence image of CDK5 staining (red) in the dorsal striatum (DS) of CDK5-NC and CDK5-KD mice. The enlarged image shows multiple mCherry-labeled neurons in the DS (scale bar = 10 μm). F Representative western blot assay of CDK5 expression in the DS. Band intensity of CDK5 relative to β-actin is shown as the mean ± SEM, n = 8 independent experiments. One-way ANOVA, Tukey’s multiple comparisons test, *p < 0.05, **p < 0.01, ***p < 0.001 vs WT