Characterization of cell type-specific CRISPR–dCas9 plasmids. (A) Schematics of the CRISPR–dCas9–KRAB–MECP2 transcriptional plasmids under the human synapsin (SYN) and GFAABC1D (GFAP) promoters that were used to target neurons and astrocytes, respectively. The GFAP–dCas9 plasmid was developed by substituting the SYN promoter. (B) Immunofluorescence of hippocampus tissue transfected with SYN–dCas9 plasmid expression in neurons. (C,D) Immunofluorescence of hippocampus tissue transfected with GFAP–dCas9 plasmid expression in astrocytes (C) but not neurons (D). NeuN stains neurons, GFAP stains astrocytes, FLAG stains for the dCas9 plasmid, and DAPI stains DNA. The dotted white box indicates the region focused on in the zoomed images.
