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Strategy for CRISPR/Cas9-mediated genome editing in ΔEx50 mice

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Presentation on theme: "Strategy for CRISPR/Cas9-mediated genome editing in ΔEx50 mice"— Presentation transcript:

1 Fig. 2. Strategy for CRISPR/Cas9-mediated genome editing in ΔEx50 mice.
Strategy for CRISPR/Cas9-mediated genome editing in ΔEx50 mice. (A) Scheme showing the CRISPR/Cas9-mediated genome editing approach to correct the reading frame in ΔEx50 mice by skipping exon 51. Gray exons are out of frame. (B) Illustration of single guide RNA (sgRNA) binding position and sequence for sgRNA-ex51. Protospacer adjacent motif (PAM) sequence for sgRNA is indicated in red. Black arrow indicates the cleavage site. Fw, foward primer; Rv, reverse primer. (C) Genomic deep-sequencing analysis of PCR amplicons generated across the exon 51 target site in 10T1/2 cells. Sequence of representative indels aligned with sgRNA sequence (indicated in blue) revealing insertions (highlighted in green) and deletions (highlighted in red). The line indicates the predicted exonic splicing enhancer (ESE) sequences located at the site of sgRNA. Black arrowhead indicates the cleavage site. (D) The muscle creatine kinase 8 (CK8e) promoter was used to express SpCas9. The U6, H1, and 7SK promoters for RNA polymerase III were used to express sgRNAs. GFP, green fluorescent protein. Leonela Amoasii et al., Sci Transl Med 2017;9:eaan8081 Published by AAAS

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