Fig. 6

Positions of guide RNAs designed for the target, here the TRPA1 promoter. Guide RNAs are numbered (compare Tables 1 and 3) and are depicted as boxes; PAM motifs are indicated by filled black squares, PAM motifs of “interfering” guide RNAs are indicated by filled grey squares. The guide RNAs yielding the highest number of calls per site (# 4 and # 9) are printed in bold. (a) This subfigure corresponds to Fig. 4 but is again shown in order to facilitate orientation in the next two subfigures. Position of guide RNAs relative to the target region on chromosome 8. Arrows mark outmost GRCh38 positions. (b) Correct position of guide RNAs enabling adapter ligation after cutting. (c) Blocking adapter binding by suboptimal positioned guide RNAs results in residual Cas9 molecules within the region, which has to be read, including the target