Work

High-efficiency multiplex biallelic heritable editing in Arabidopsis using an RNA virus

Year: 2022

Type: article

Abstract: 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 tRNA Ileu that induces efficient multiplex somatic and biallelic heritable editing in Arabidopsis . Inclusion of tRNA Ileu enhances the systemic movement of TRV and the mutant phenotype is visible in the initial TRV::sgRNA-tRNA Ileu 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::tRNA Ileu system described here allows generation of biallelic edited plants in a single generation and is amenable for large-scale high throughput CRISPR screens.

Source: PLANT PHYSIOLOGY

DOI: 10.1093/plphys/kiac159

Authors: Ugrappa Nagalakshmi, Nathan Meier, Jau-Yi Liu, Daniel F Voytas, Savithramma P Dinesh-Kumar

Institutions: Innovative Genomics Institute, University of California, Davis, University of California, Berkeley, University of Minnesota

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Zero hunger

Open Access status: hybrid

Estimated APC paid: $3,411