RPS5A Promoter-Driven Cas9 Produces Heritable Virus-Induced Genome Editing in Nicotiana attenuata

Abstract

The virus-induced genome editing (VIGE) system aims to induce targeted mutations in seeds without requiring any tissue culture. Here, we show that tobacco rattle virus (TRV) harboring guide RNA (gRNA) edits germ cells in a wild tobacco, Nicotiana attenuata, that expresses Streptococcus transgenic plants expressing SpCas9 under the control of 35S promoter and infected rosette leaves with TRV carrying gRNA. Gene-edited seeds were not found in the progeny of protein S5 A (RPS5A) promoter fused to SpCas9 was employed to induce the heritable gene editing with TRV. monoallelic mutations at three target genes in N. attenuata seeds with TRV-delivered guide RNA. These monoallelic progenies. This editing method provides an alternative way to increase the heritable editing efficacy of VIGE.