Multiplexed heritable gene editing using RNA viruses and mobile single guide RNAs

An in planta gene editing approach was developed wherein Cas9 transgenic plants are infected with an RNA virus that expresses single guide RNAs (sgRNAs). The sgRNAs are augmented with sequences that promote cell-to-cell mobility. Mutant progeny are recovered in the next generation at frequencies ranging from 65 to 100%; up to 30% of progeny derived from plants infected with a virus expressing three sgRNAs have mutations in all three targeted loci. An efficient and multiplexed in planta gene editing approach is developed by infecting Cas9 transgenic plants with an RNA virus that expresses single guide RNAs carrying sequences that confer cell-to-cell mobility.