Fig. 5: A22p peptide-assisted delivery of SpyCas9 for gene editing at disease-relevant genomic sites.

a Schematic of the target sequences of Cas9 RNPs and the protospacer adjacent motif (PAM) for TH and mGluR5 knockout. b In vitro knock-out of the TH and mGluR5 genes based on direct delivery of the Cas9 RNPs. Indels quantified by NGS. n = 4 for each group, data are presented as mean values with individual data points. c In vivo knock-out of the TH and mGluR5 genes based on intraparenchymal injections of the Cas9 RNPs into mouse brains. RNPs assembled with a 1.5:1 mole ratio of sgRNA to Cas9 protein. Editing efficiency at the DNA level (indel throughout the whole striatum) quantified by NGS. n = 4 for the non-targeting control group and n = 6 for the experimental group. Data are presented in box plots where the lower bound of the lower whisker shows the minimum, the lower bound of the box shows the lower quartile, the center of the box shows the median, the upper bound of the box shows the upper quartile and the upper bound of the upper whisker shows the maximum. d Editing efficiency at the mRNA level (throughout the whole striatum) quantified by qPCR. n = 4 for the non-targeting control group and n = 5 for the experimental group, data are presented as median values ±s.e.m. Statistical analyses (unpaired t test) were performed for (c, d), and p values were indicated with each set of quantification. Images in (b, c) were created with biorender.com.