An optimised CRISPR Cas9 and Cas12a mutagenesis toolkit for Barley and Wheat
Based on the results of our testing, we describe and provide a GoldenGate modular cloning system for Cas9 and Cas12a use in barley and wheat. Proven Cas nuclease and guide expression cassette options found in the toolkit will facilitate highly efficient simplex and multiplex mutagenesis in both species. We incorporate GRF-GIF transformation boosting cassettes in wheat options to maximise workflow efficiency.
Authors
- Wendy Harwood (ORCID: https://orcid.org/0000-0002-4323-9009)
- Tom Lawrenson
- Martha Clarke
- James K. M. Brown (ORCID: https://orcid.org/0000-0001-8719-2268)
- Burkhard Steuernagel (ORCID: https://orcid.org/0000-0002-8284-7728)
- Macarena Forner
- Rachel Kirby
Institutions
- John Innes Centre (GB)
- Norwich Research Park (GB)
Publication Details
- Journal
- Plant Methods
- Published
- 2024-08-13
- DOI
- https://doi.org/10.1186/s13007-024-01234-y
- Citations
- 10
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 2.34
Funders
- Bill and Melinda Gates Foundation
- Foreign, Commonwealth and Development Office
- Directorate for Biological Sciences
- Biotechnology and Biological Sciences Research Council