Figure 2: Overview of the Digenome-seq Workflow after ABE-Mediated Single Letter Substitution (IMAGE)

Institute for Basic Science

Figure 2: Overview of the Digenome-seq Workflow after ABE-Mediated Single Letter Substitution

Caption

ABE7.10 (light blue, with guide RNA represented as white filament) induces the conversion of adenine (A) to inosine (I) at the target DNA site. Since I is read as G during DNA replication, this leads to the conversion of A to G. In addition, the complex also causes a DNA cut in one strand. As the Digenome-seq technique works by detecting double-strand breaks, the researchers used a protein known as Endo V to produce a cut in the other DNA strand, nearby the I. At the end of the Digenome-seq procedure, shown in the circular Circos plot, it is possible to evaluate and count the on-target (red arrow) and off-target (black bars) positions. The same results were obtained with hAAG and Endo VIII, in place of Endo V, showing Digenome-seq reproducibility and reliability.

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