Plasmid-Free Genome Editing of Cabbage and Chinese Cabbage

CRISPR-Cas9-mediated gene editing is enabled by the same methods as the production of genetically modified organisms. The only difference is that the transferred DNA sequence is sorted out of the plant at the end of the experiment, making the plant transgene-free. However, researcher Roman Jerala of National Institute of Chemistry in Slovenia and colleagues say that the insertion of the transgene at the earlier part of the experiment can cause unwanted mutations. Also, further expression of the transgene in the plant can cause off-target mutations. Therefore, the team proposed a method to skip transgene production in gene editing.

The team transformed Cas9 enzymes and guide RNA sequence with ribonucleoprotein into plant protoplasts using PEG-mediated transformation. Results showed up to 25 percent mutation frequency in genes FRI and PDS in the said species. They also observed positive correlation between amount of CRISPR components and mutation rate. The team aims to target other genes and study edited protoplast regeneration in the future.

For more information, read the article in Frontiers in Plant Science.