Figure 2
From: Efficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway
diRNAs are not detected at endogenous genes in Arabidopsis and rice. (A) Three independent CRISPR/Cas9 constructs target to AtBRI1. The top panel is a schematic representation of AtBRI1 gene and three CRISPR/Cas9 gRNA target sites. Northern blot analysis was performed with LNA probes (see Supplemental Table 1) in AtBRI1 gene targeted CRISPR/Cas9 T1 transgenic lines. An miR167 probe was used as a control. (B) Northern blot analysis of small RNAs for additional endogenous genes (GAI, AtGL2, At5g36250, At2g36490 and At5g04560) targeted by CRISPR/Cas9. The Col0 accession and T1 transgenic plants were analyzed. (C) Detection of antisense transcripts at GAI by qRT-PCR, after targeting by CRIPR/Cas9. The same transgenic plants were used as in Fig. 2B. GAI-as indicates GAI antisense transcript. IGN33 was analyzed as a control. Black bar; Col0 with RT, gray bar; GAI targeting CRIPR/Cas9 with RT, white bar; without RT control. N = 3. N.D.; not detected. (D,E) Detection of small RNAs in endogenous genes targeted by CRISPR/Cas9 (D) and TALEN (E) in T0 transgenic rice. Horizontal line indicates TALEN target site and probe. U6 and miR159 were probed as controls. (A,B,D,E) Northern blotting image was cropped nearby signals.
