CRISPR-Cas9 Genome Editing in Rapeseed

CRISPR-Cas9 is a valuable tool for research and has been widely applied to different plant species. However, the efficiency of this method is not available for the allotetraploid rapeseed (Brassica napus), an important oilseed crop. Huazhong Agricultural University researchers evaluated the mutation efficiency of the CRISPR-Cas9 for 12 genes in rapeseed, including paralogous genes.

CRISPR efficiently induced targeted mutations both for single locus and multiple loci targeting in rapeseed. CRISPR also induced mutations in paralogous genes, or different genes with the same function in a species. Since the paralogous genes in rapeseed have highly similar sequences, they were successfully targeted using only a single sgRNA. No off-target mutations were identified in the CRISPR-Cas9 lines, indicating that the mutations induced were highly specific.

Half of the mutations, including homozygotes, bi-alleles, and heterozygotes were also stably inherited in the next generation (T1) without any new mutations or reversions.These results demonstrate that CRISPR-Cas9 can be an efficient tool for targeted genome modifications at multiple loci in rapeseed.

For more information, read the article in Scientific Reports.