Differential editing efficiencies in cereal crops: a comparative analysis of tRNA and ribozyme multiplexed guide delivery

• DOI: 10.3389/fpls.2024.1426184
• Publication Date: 2024-12-05T16:12:51Z
• Authors: Matthew J. Milner; Manisha Sharma; Ruth E. Bates; Michelle Whiting; Melanie S. Craze; Peter Miller; Jack Brooks; Allan Kouidri; Emma J. Wallington
• Keywords: Guide RNA

Cereal transformation and gene editing can be a complex costly undertaking. It is therefore important to validate understand the performance of components achieve high rates editing. Here, we have made direct comparison different CRISPR/Cas9 guide systems target genome in three cereal species. We show that sequences driven by same pol II promoter rice, wheat barley large differences efficiency. The seen were based on way guides presented factors outside sequence itself. While both tRNA system ribozyme performed well their effectiveness varied barley. Specifically, outperformed system, achieving higher stable transformed plants. Overall, levels are observed all species when strong expression SpCas9 coupled with CmYLCV drive array RNAs. Stable inheritance also achievable plants sampled shortly after tissue culture concludes. above 85% species, particularly mutations detected early emerge from culture.