Fig. 6

Specific virus targeting via CasRx with no observed collateral activity. a Schematic of CasRx targeting of TRBO-BFP and PVX-GFP viruses in synchronous co-infected leaves with the two different viral vectors, and the possible outcomes. b BFP monitoring to assess the Cas13-mediated virus interference activities in Agro-infiltrated wild type N. benthamiana leaves in transient assays (left). Images were taken 3 days post-infiltration. NS, non-specific crRNA; Rep, replicase. RT-qPCR analysis of TRBO-BFP knockdown with Rep-T3 crRNA used against TRBO-BFP virus (right). The transcript levels of TRBO-BFP are shown relative to the transcript level of TRBO-BFP virus targeted with (NS) crRNA. Values shown as mean ± SEM (n = 3). c BFP and GFP monitoring to assess the Cas13-mediated virus interference activities in Agro-infiltrated wild type N. benthamiana leaves in transient assays (left). Images were taken 3 days post-infiltration. NS, non-specific crRNA; Rep, replicase. RT-qPCR analysis of TRBO-BFP and PVX-GFP knockdown with Rep-T3 crRNA (right). The transcript levels of TRBO-BFP are shown relative to the transcript level of the non-targeted virus (PVX-GFP), and the transcript level of both viruses is shown relative to the transcript level of both viruses targeted with (NS) crRNA. Values shown as mean ± SEM (n = 3). d GFP monitoring to assess the Cas13-mediated virus interference activities in Agro-infiltrated wild type N. benthamiana leaves in transient assays (left). Images were taken 3 days post-infiltration. NS, non-specific crRNA. RT-qPCR analysis of PVX-GFP knockdown with GFP-T2 crRNA used against PVX-GFP virus (right). The transcript levels of PVX-GFP are shown relative to the transcript level of PVX-GFP virus targeted with (NS) crRNA. Values shown as mean ± SEM (n = 3). e BFP and GFP monitoring to assess the Cas13-mediated virus interference activities in Agro-infiltrated wild type N. benthamiana leaves in transient assays (left). Images were taken 3 days post-infiltration. NS, non-specific crRNA. RT-qPCR analysis of TRBO-BFP and PVX-GFP knockdown with GFP-T2 crRNA (right). The transcript levels of PVX-GFP are shown relative to the transcript level of the non-targeted virus (TRBO-BFP), and the transcript level of both viruses is shown relative to the transcript level of both viruses targeted with (NS) crRNA. Values shown as mean ± SEM (n = 3)