Fig. 3

Application of Nme2ABE8e-797 to disease-relevant loci in Neuro-2a cells and mice. a Schematic representation of the strategy that restores the in-frame deletion of 150 nt (LMNA Δ150) of the LMNA transcript (left). The target sequence for reverting the LMNA c.1824 C > T mutation in mouse Neuro-2a LMNA c.1824 C > T mutation cell lines is shown. The PAM-targeted and sgRNA-targeted sequences are shown in green and black, respectively. The substituted bases are indicated in red (right). b Representative Sanger sequencing chromatograms of the LMNA locus. The red arrow indicates the substituted nucleotide. c Comparison of A-to-G editing efficiency produced by Nme2ABE8e and Nme2ABE8e-797 using Sanger sequencing in mouse Neuro-2a LMNA c.1824 C > T mutation cell lines. Bars represent mean values, and error bars represent the s.d. of three independent biological replicates, with * and ** representing P < 0.05 and 0.01, respectively (two-tailed unpaired t test). d RT‒PCR analysis of mis-spliced LMNA mRNA. The red arrow indicates the mis-spliced LMNA mRNA. e The target sequence at the TYR (p.D42G) locus. The PAM-targeted and sgRNA-targeted sequences are shown in green and black, respectively. The substituted bases are marked in red. f Characterization of the targeted modifications in TYR (p.D42G) mice using next-generation sequencing. g Analysis of off-target effects in TYR mutant mice using next-generation sequencing. Data are presented as mean ± s.d. (n = 2 mice)