Extended Data Fig. 7: TRV is not transmitted to the next generation.
From: Multiplexed heritable gene editing using RNA viruses and mobile single guide RNAs
RT-PCR was performed using total RNA extracted from progeny heterozygous for mutations at PDS of plants infected with TRV editing vectors. a, Progeny RNA from a parental plant infected with TRV-sgNbPDS-tRNAMet (lanes 1–2) and TRV-sgNbPDS-tRNAIleu (lanes 3–4), TRV-infected control (lane 5) and mock-infected control plant tissue (lane 6) using primers that amplify the TRV 3’-end (upper panel) and endogenous NbPP2A gene (bottom panel). Lane 7, TRV plasmid template used in PCR as a positive control. b, Progeny RNA from parental plant replicate 1 infected with TRV-sgNbPDS-FT (lanes 10–17), TRV-infected control (lanes 9, 19, and 29) and water control (lanes 8, 18, and 28) using primers that amplify TRV across the sgNbPDS-FT insert (upper panel) and endogenous NbEF1a gene (bottom panel). c, Progeny RNA from parental plant replicate 2 infected with TRV-sgNbPDS-FT (lanes 20–27). d, Progeny RNA from parental plant replicate 3 infected with TRV-sgNbPDS-FT (lanes 30–37). Lanes M1, 100 bp DNA ladder; M2, 1 kb DNA ladder; M3, NEB 1 kb+ ladder. Conclusions were the same for experiments performed at different institutions: the experiment in panel a was performed at UC Davis; the experiments in panels b–d were performed at the University of Minnesota.
