Journal
MOLECULAR PLANT PATHOLOGY
Volume 20, Issue 10, Pages 1463-1474Publisher
WILEY
DOI: 10.1111/mpp.12849
Keywords
Barley stripe mosaic virus; CRISPR mutagenesis; delivery; gRNA; maize; Nicotiana benthamiana; targeted mutagenesis; wheat
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Funding
- National Key R&D Program of China [2016YFD0100502]
- Transgenic Research Program of China [2016ZX08010-001]
- Project for Extramural Scientists of SKLAB [2019SKLAB1-14]
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Plant RNA virus-based guide RNA (gRNA) delivery has substantial advantages compared to that of the conventional constitutive promoter-driven expression due to the rapid and robust amplification of gRNAs during virus replication and movement. To date, virus-induced genome editing tools have not been developed for wheat and maize. In this study, we engineered a barley stripe mosaic virus (BSMV)-based gRNA delivery system for clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-mediated targeted mutagenesis in wheat and maize. BSMV-based delivery of single gRNAs for targeted mutagenesis was first validated in Nicotiana benthamiana. To extend this work, we transformed wheat and maize with the Cas9 nuclease gene and selected the wheat TaGASR7 and maize ZmTMS5 genes as targets to assess the feasibility and efficiency of BSMV-mediated mutagenesis. Positive targeted mutagenesis of the TaGASR7 and ZmTMS5 genes was achieved for wheat and maize with efficiencies of up to 78% and 48%. Our results provide a useful tool for fast and efficient delivery of gRNAs into economically important crops.
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