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Fig. 5 | Genome Biology

Fig. 5

From: CRISPR-Cas13d mediates robust RNA virus interference in plants

Fig. 5

CasRx catalytic activity is required for RNA virus interference. a Illustration of the targeting activity of the catalytically active and catalytically deactivated (dCasRx) CasRx variants against the targeted TRBO-GFP virus. b 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. 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 and anti-HA antibodies. α-GFP, anti-GFP antibody, α-HA, anti-HA antibody. Ponceau staining served as loading control. d RT-qPCR analysis of TRBO-GFP knockdown with the catalytically active and catalytically deactivated CasRx variants. For each CasRx 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)

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