Fig. 2

TRBO-GFP-based screening of different Cas13 orthologues for efficient virus interference. a GFP monitoring to assess the Cas13-mediated virus interference activities in Agro-infiltrated wild type N. benthamiana leaves in transient assays. Images were taken 3 days post-infiltration. NS, non-specific crRNA. b Relative fluorescence intensity quantification of leaf images. For each Cas13 variant, GFP signal intensity of each targeting crRNA (GFP-T1 and GFP-T2) is shown relative to the non-targeting (NS) crRNA. Values shown as mean ± SEM (n = 3). c Western blot analysis of the abundance of the virus expressed GFP protein to confirm the Cas13-mediated TRBO-GFP virus interference. Protein blots were developed with anti-GFP antibody. α-GFP, anti-GFP antibody. Ponceau staining served as loading control. d Quantification of the western blot data. For each Cas13 variant, the abundance of the GFP protein with the targeting crRNAs (GFP-T1 and GFP-T2) is shown relative to the non-targeting (NS) crRNA. Error bars indicate SEM (n = 3). e RT-qPCR analysis of TRBO-GFP knockdown with different Cas13 variants using the two position-matched crRNAs. For each Cas13 variant, knockdown efficiency of each targeting crRNA (GFP-T1 and GFP-T2) is shown relative to the non-targeting (NS) crRNA. Values shown as mean ± SEM (n = 3)