Fig. 2.

Editing efficiency analysis on 14 HCT_gRNAs. a Location of sgRNA target sites in the HCT gene. The open reading frame of the HCT gene is represented with an orange arrow. The binding site and direction of individual HCT_gRNA are represented with green arrows. b The relative efficiency of individual HCT_gRNA is expressed as GFP:RFP ratios. NC: negative control. Data represent 5–7 biological replicates. For details of the individual sgRNAs, see Additional file 1. Asterisks indicate significant differences compared to NC (Student’s t-test; *P < 0.05; **P < 0.01; ***P < 0.001). c Confirmation of mutations at the HCT_gRNA12 and HCT_gRNA14 target sites. Sequences spanning the target sites on HCT transcript were amplified and analyzed by Sanger sequencing. The unmodified (WT) sequence of the corresponding region is shown on top of the chromatogram with the spacer sequences in bold and the PAM sequences underlined