RT Journal Article SR Electronic T1 High efficiency multiplex biallelic heritable editing in Arabidopsis using an RNA virus JF bioRxiv FD Cold Spring Harbor Laboratory SP 2022.01.20.477144 DO 10.1101/2022.01.20.477144 A1 Nagalakshmi, Ugrappa A1 Meier, Nathan A1 Liu, Jau-Yi A1 Voytas, Daniel F. A1 Dinesh-Kumar, Savithramma P YR 2022 UL http://biorxiv.org/content/early/2022/01/22/2022.01.20.477144.abstract AB Delivery of gene editing components such as the Cas nuclease and single guide RNAs (sgRNAs) into plant cells is commonly accomplished by agrobacterium-mediated transformation. Although Arabidopsis is easy to transform, generation of biallelic edited plants requires screening a large number of plants in subsequent generations. Here, we describe optimization of the Tobacco rattle virus (TRV) for in planta delivery of sgRNAs fused to a tRNAIleu that induces efficient multiplex somatic and biallelic heritable editing in Arabidopsis. Inclusion of tRNAIleu enhances the systemic movement of TRV and the mutant phenotype is visible in the initial TRV::sgRNA-tRNAIleu infected Arabidopsis, which allows for the uncovering of lethal phenotypes. Mutant progeny are recovered in the next generation (M1) at frequencies ranging from 30-60%, with 100% mutant recovery in the following (M2) generation. TRV::tRNAIleu system described here allows generation of biallelic edited plants in a single generation and is amenable for large-scale high throughput CRISPR screens.Competing Interest StatementThe authors have declared no competing interest.