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. 2016 Feb 26;7:10770. doi: 10.1038/ncomms10770

Figure 6. Chromosomal rearrangements in cancers induced by CRISPR/Cas9-multiplexing.

Figure 6

(a) CRISPR/Cas9-induced intra-chromosomal deletion of an 18 Kb fragment by combinatorial sgRNA targeting. PCR screening for all possible intra-chromosomal fusions on chromosomes with more than one sgRNA target site was performed across pancreatic cancers. The gel image shows respective PCRs at the Cdkn2a locus. sgRNA target sites in Cdkn2a-ex1ß/ex2 and PCR primers are indicated by red arrow heads and black arrows, respectively. Sanger sequencing confirmed the ex1β-ex2 fusion. del, deletion; ex, exon. (b,c) M-FISH of cell lines revealed unbalanced inter-chromosomal translocations induced by combinatorial CRISPR/Cas9 targeting in two out of six cancers. Fusion products were detected by PCR and sequencing. Arrows indicate primer positions. del, deletion; der, derivative; t, translocation.