Figure 4.

STRIDE for sequence detection on dsDNA: (A) CRISPR/Cas9-mediated induction of double-strand breaks (DSB); (B) Modified deoxynucleotides are enzymatically conjugated to 3′ DNA ends by Pol I; (C) Two different primary antibodies, directed against the modified nucleotides bind in close proximity; (D) Secondary antibodies conjugated with oligonucleotides bind to primary antibodies; (E) Another oligonucleotide binds to the two oligonucleotides from D., forming a circular DNA template; (F) One oligonucleotide from D. acts as primer for elongation in a rolling circle mechanism by a DNA polymerase using the circular DNA from E. as template thereby generating a ssDNA with repetitive motifs; (G) Hybridization of fluorophore-labelled oligonucleotides to the ssDNA from the rolling circle. More information on the graphical elements is given in the box within the figure.