Fig. 7: Multiplexing of fusion-positive samples with varying breakpoints and fusion partners. | Nature Communications

Fig. 7: Multiplexing of fusion-positive samples with varying breakpoints and fusion partners.

From: Partner independent fusion gene detection by multiplexed CRISPR-Cas9 enrichment and long read nanopore sequencing

Fig. 7

a Schematic overview of the multiplexing approach. Samples are prepared and subjected to the Cas9-enrichment individually and pooled equally pre-sequencing. The library-pool is sequenced on a single ONT flow cell and NanoFG detected fusion genes and designed fusion-specific breakpoint primers. Original samples are subjected to breakpoint PCR to identify the sample-of-origin for each fusion gene. b Coverage plots for KMT2A (targeted). Dotted lines (green) indicate the crRNA-directed Cas9 cleavage positions and dashed lines (red) indicate breakpoint positions. Arrows indicate the directionality of reads created from the specific crRNA design. c Breakpoint locations within the KMT2A gene for the different identified fusion genes. Breaks cluster between Exon 8 and 12 and reciprocal fusion genes are highlighted in the same color. d Enrichment across the fusion junctions. Source data are provided as a Source Data file.

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