Fig. 5: PIS-mediated large deletions.
From: Efficacy and long-term safety of CRISPR/Cas9 genome editing in the SOD1-linked mouse models of ALS
a Schematic representation of the cloning and enzymatic analysis of a 6.6 kb fragment from hSOD1 in G1H/Cas9 mice. The restriction enzyme sites used for analysis are labeled. Red arrowhead indicates the Cas9 cleavage site. Red arrows indicate plasmid vector DNA. Six types of clones were identified. Type 1 was Wt-like, the other five types showed smaller inserts. b–m Twelve deletions were identified, ranging from 325 to 4608 bp in size. Sequencing chromatogram of the DNA sequence around the junction is shown on the top. The PIS is color coded in red. The 5’ junction site is indicated by an arrow. Description of the deletions is based on the NCBI reference sequence (NC_000021.9) for human SOD1. n Schematic representation of deletions showing the sizes and locations (diagrams are not drawn to scale for clarity). Six Alu elements are shown by filled boxes, and the deleted regions are shown by lines. Deletions mediated by different PIS in the same Alu elements are shown on the top of the lines. The total number of deletions are summarized on the right, and detailed in Table 1.
