Studies have been made to optimize the CRISPR/Cas9 system for crop breeding.
The main reasons for these studies are the avoidance of transgene
integration and reduction of off-target mutations. Zhen Liang of the Chinese
Academy of Sciences aimed to develop an efficient genome editing method for
bread wheat using CRISPR/Cas9 ribonucleoproteins (RNPs).

The developed protocol takes only seven to nine weeks. Four to five
independent mutants were produced from 100 immature wheat embryos. Further
analysis revealed much lower chance of off-target mutations in RNP-mediated
genome editing than with CRISPR/Cas9 DNA. Furthermore, no off-target
mutations were detected in the mutant plants.

Since no foreign DNA is used in CRISPR/Cas9 RNP-mediated genome editing, the
mutants obtained were completely transgene-free. This method could be widely
applicable for producing genome-edited crops.





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