Figure 3
From: Efficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway
diRNAs are not detected at endogenous repeat regions. (A–C) The repeat regions and the targeted T1 sequences are shown for the (A) At1g31290 repeat1, (B) At1g31290 repeat2, and (C) At5g54700, respectively. The top gray and black horizontal bars represent the repeats, and the CRISPR/Cas9 target sites are indicated by arrows. The sequence of the repeats is below the gray/black schematic. Below the reference repeat sequence are the sequencing results from independent T1 plants. See also Supplemental Figs 4 and 5. (D–F) Northern blot analysis for small RNAs in the indicated Col0 control, T1, T2 with CRISPR/Cas9 transgene (T2 w) or without CRISPR/Cas9 transgene (T2 wo). (D) At1g31290 repeat1, (E) At1g31290 repeat2, and (F) At5g54700, respectively. miR167 was probed as control. Northern blotting image was cropped nearby signals. (G) qRT-PCR detection of antisense transcripts in At1g31290 repeats targeted by CRISPR/Cas9. At1g31290-as indicates At1g31290 antisense transcript. IGN33 was analyzed as control. Black bar; Col0 with RT, gray bar; At1g31290 targeting CRIPR/Cas9 with RT, white bar; without RT control. N = 3. N.D.; not detected.
