Figure 5: Mutations of the ASPM Gene in Monkey Tissues.
From: Promoting Cas9 degradation reduces mosaic mutations in non-human primate embryos
(A) T7E1 digestion of the targeted ASPM gene in embryos injected with WT-Cas9 or Ubi-Cas9. Green arrows indicate the mutations in single cells isolated from 2-cell or 4-cell embryos. (B) ASPM immunofluorescent staining of 2-cell-stage monkey embryos showing that injection of Ubi-Cas9 mRNA and gRNAs for targeting ASPM can deplete the spindle localization (arrows) of ASPM. Scale bar: 20 μm. (C) Sequence analysis of tissue DNAs from a live monkey indicating mutations in the ASPM gene that was targeted by two gRNAs for exon 3 and exon 10. The numbers of cloned DNA for sequencing and mutated DNA clones are indicated in parentheses (mutated clones/total clones). Targeted sequence regions are indicated in red.
