High-Efficiency, Heritable Gene Editing Using RNA Viruses

Gene editing with sequence specific nucleases offers great promise for advancing basic and applied plant research. Nearly all heritable mutations created from gene editing reagents, however, rely on tissue-culture to regenerate edited plant cells. Tissue-culture works on limited genotypes, is very time consuming, and is often technically challenging which dramatically limits the scope and throughput of gene editing in plants. One potential solution is through the use of RNA viral vectors to deliver CRISPR reagents. Up until this point this approach has been limited by low frequency somatic gene editing and the inability to efficiently generate heritable indels. Through the use of augmented sgRNAs expressed from RNA Viral Vectors we have demonstrated high frequency somatic and heritable tissue-culture free gene editing in Nicotiana benthamiana. We anticipate this method will be broadly applicable and considerably increase the feasibility of plant gene editing.