Enhanced Green Fluorescent Marker Effective in Transforming Grapes Using CRISPR

CRISPR-Cas9-mediated genome editing in grapes was first reported in 2016. With the increasing interest in using this new breeding technique in grapes, scientists from the Chinese Academy of Sciences evaluated genome editing in grapes using newly developed CRISPR-based tools such as the CRISPR/LbCpf1 system to increase existing evidence that gene editing tools are useful in improving the crop.

The scientists evaluated four different markers and found that the Enhanced Green Fluorescent Protein  (EGFP) was not harmful to grape cells, but its signal was not easily detected. Scientists then used EGFP and RUBY, another marker, to help screen transformed grape cells based on fluorescence and red betalain. They then tested the activities of CRISPR/LbCpf1, CRISPR-Cas9, and cytidine base editor. Further examination revealed that using the CRISPR/LbCpf1 with the aid of the EGFP marker is effective in generating targeted mutagenesis in grapes and is capable of editing multiple targets simultaneously using only a single crRNA array. The scientists also noted that elevated temperature could improve the activity of the CRISPR/LbCpf1 in grapes.

The study provides evidence that can be used for further gene editing research on grapes.

Read the pre-print of the publication in bioRxiv.