Figure 1: Cas9-induced DNA breaks promote interchromosomal translocations.
From: Targeted genomic rearrangements using CRISPR/Cas technology
(a) Schematic depicting the overall strategy for generating chromosomal rearrangements. Cas9 from S. pyogenes (SpCas9) is co-expressed with two single-guide RNAs (sgRNA 1 and 2) that direct DNA cleavage at each targeted genomic site. (b) The CD74-ROS1 rearrangement results from a translocation between chromosomes 5 and 6. Shown are the intronic sites where Cas9 was targeted. (c,d) PCR detection of the (c) CD74-ROS1 Der(6) and (d) ROS1-CD74 Der(5) genomic breakpoint junctions from HEK 293T cells in which Cas9 was expressed with no sgRNA (vector), CD74 sgRNA alone, ROS1 sgRNA alone, or both CD74 and ROS1 sgRNAs. (e) Sequence chromatogram of the detected CD74-ROS1 fusion transcript from cells in which Cas9 and both CD74 and ROS1 sgRNAs were expressed. Data shown are representative results from a total of three independent experiments.
