Supplementary Figure 4: Analysis of the diversity of editing outcomes at PigA locus. | Nature Biotechnology

Supplementary Figure 4: Analysis of the diversity of editing outcomes at PigA locus.

From: Repair of double-strand breaks induced by CRISPR–Cas9 leads to large deletions and complex rearrangements

Supplementary Figure 4

PigA locus was edited using 5’ intronic guide (#15) in biological quadruplicate. Duplicate PCR reactions spanning the cut site performed on DNA extracted from the bulk of PigA negative cells and resolved on an agarose gel (product size 3500bp, 1F/1R primer pair, Fig. S2A). (A) Experimental considerations. (B) Deletion fingerprints. Cell line: JM8 – the original mouse embryonic stem cell line (transfected with gRNA and Cas9 PiggyBac vectors); CAS – subclone of JM8 expressing Cas9 from a single-copy lentiviral transgene (transfected with only gRNA PiggyBac vector). Ladder scale is in kilobases.

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