Extended Data Fig. 8: Immunohistochemistry of SpCas9 and NeuN i in the Mef2c L35P+/- mice and evaluation of Off-targeting effects by AeCBE in vivo.
From: Whole-brain in vivo base editing reverses behavioral changes in Mef2c-mutant mice
(a) Immunohistochemistry of SpCas9 and NeuN in the prefrontal cortex of AAV-AeCBE injected mice. (b) Quantification of SpCas9 and NeuN double-positive neurons in the total NeuN cells (n = 7 slices from 3 mice). (c) Immunostaining of SpCas9 and NeuN in the hippocampus of AAV-AeCBE injected mice. (d) Quantification of SpCas9 and NeuN double-positive neurons in the total NeuN cells (n = 12 slices from 4 mice). (e) Immunohistochemistry images of Mef2c and NeuN in the hippocampus of AAV-AeCBE injected mice. (f) Quantification of Mef2c and NeuN double-positive neurons in the total NeuN cells (n = 12 slices from 4 mice). (g) Off-target sites detected by GUIDE-seq, with sgRNA targeted to Mef2c L35P region. More reads indicated more likely to be targeted by SgC8 and SpG-Cas9. (h) Venn diagrams of off-target sites detected by the in-silico method and the GUIDE-seq method. (i) Off-target editing analysis at all 11 candidate sites by next-generation sequencing in vivo. (n = 3 mice, OT2-C7: P < 0.0001; OT4-C8: P = 0.0214; OT5-C8: P = 0.0243). Statistical values represent the mean ± s.e.m. * P < 0.05, *** P < 0.001, unpaired two-tailed Student’s t-test.
