Fig. 2: Cleavage of the ITS region of the 45S rDNA repeats in Arabidopsis using SaCas9.
From: Using CRISPR-Kill for organ specific cell elimination by cleavage of tandem repeats
a Cleavage of the conserved ITS region of the 45S rDNA using SaCas9 in the wild type resulted in a dramatic decrease of the number of transformants in all three ITS-targeting lines (Ubi-SaCas9-ITS1-1, Ubi-SaCas9-ITS1-2, and Ubi-SaCas9-ITS2-1) compared to the control line, Ubi-SaCas9-ADH1 (n = 3 biologically independent experiments). Data are presented as normalized mean values ± SD. P = 0.00043 for ITS1-1; P = 0.00085 for ITS1-2; P < 0.0001 for ITS2-1. b The survival rate of lines transformed with a catalytically inactive SaCas9 (dCas9) targeting ADH1 (Ubi-dCas9-ADH1) and ITS2 (Ubi-dCas9-ITS2-1) was analyzed. No reduction in survival rate could be observed in the ITS2-1 line compared to the ADH1 line (n = 3 biologically independent experiments). Data are presented as normalized mean values ± SD. c SaCas9 (blue) is under the expression of an exchangeable promoter (light pink). Utilization of SaCas9 under expression of a constitutive results in cleavage of the 45S rDNA in all cells (red), leading to cell death. By exchanging the constitutive promoter with a tissue-specific promoter, it is possible to obtain plants devoid of individual organs. Statistical differences were calculated using the two-tailed t-test with unequal variances: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Source data are provided as a Source data file.
