Fig. 4
From: CRISPR guides induce gene silencing in plants in the absence of Cas
Guide mismatches at position 10 and 11 abolish GIGS, indicating AGO dependence. a Illustration of multi-guide expression from TRV targeting the PDS transcript. For each of the 28-nt guides (crRNA1, crRNA2, crRNA 3) a variant m-guide 1 was designed. For mg 1[mm5,6], each crRNA contained two base pair mismatches at positions 5,6, for mg 1[mm10,11] mismatches at positions 10,11, and mg 1[mm21,22] contained mismatches at positions 21,22. b Representative images of leaves following TRV systemic delivery of m-guide 1 targeting PDS, in addition to the three variants of m-guide 1. TRV expressing a non-targeting guide (NT-guide) and TRV with a region of antisense sequence to PDS (asPDS) served as controls. c SPAD meter readings from photobleached (loss of green color) leaf samples. Data collected from a total of six independent leaves from two experiments. d Quantification of the PDS transcript using qPCR for the same samples as measured in c. Data standardized to an endogenous transcript and normalized to TRV expressing NT-guide. Statistical comparisons were made between the NT-guide and each treatment using a one-sided Mann-Whitney U test with Benjamini-Hochberg (BH) multiple testing correction. Samples with p values less than 0.05 (*) and 0.01 (**) are indicated. n.s., non-significant difference (p > 0.05)