Figure 1
From: Efficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway
diRNAs are generated from transgenes targeted by CRISPR/Cas9. (A) A schematic representation of the 35S::GU-US reporter system. The GU-US reporter contains a CRISPR/Cas9 gRNA target site located within the direct U repeats of GU-US gene. (B) Representative GUS staining images of plants containing the GU-US transgene, in the presence or absence of the CRISPR/Cas9 system. The transgenic line that does not carry CRISPR/Cas9 was used as negative control. (C) The relative repair rate determined by qPCR. The primers for qPCR amplify the CRISPR/Cas9 target linker sequence and S part of GUS gene, and the repair rate was calculated. Error bars indicate standard error of 3 repeats. (D) Detection of small RNAs by Northern blot. The U part of GUS gene was used for the GUS probe. A miR167 probe was used as a loading control. Northern blotting image was cropped nearby signals. (E) Distribution of small RNA on 35S::GU-US transgene. Two independent T1 transgenic lines were analyzed. The y axis represents the number of 21 nt long small RNA reads within 100 bp sliding windows with a step size of 1 bp, numbers in (+) and (−) values represent the reads of small RNAs derived from sense and antisense strands, respectively. (F) Detection of small RNAs by Northern blot in 35S::GU-US transgenic rice T0 generation. The U part of GUS gene was used for GUS probe. The U6 probe was used as a loading control. Northern blotting image was cropped nearby signals.
