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Crop Biotech Update

Genetic Editing of Grapevine and Apple Protoplasts using CRISPR/Cas9 Ribonucleoproteins

December 7, 2016

The availability of genome sequences and genome editing tools is set to revolutionize the field of fruit biotechnology through the introduction of targeted genetic changes. While plasmid-mediated delivery of genome editing components is very efficient, it presents some drawbacks. Furthermore, it may be cut-off by current process-based GMO regulations, complicating its path to commercialization.

Mickael Malnoy of the Foundation Edmund Mach in Italy, together with colleagues from various institutions explored the direct delivery of purified CRISPR/Cas9 ribonucleoproteins (RNPs) to the protoplast of grape cultivar Chardonnay, and apple cultivar, such as Golden delicious, for efficient targeted mutagenesis.

The team targeted MLO-7, a susceptible gene, in order to increase resistance to powdery mildew (PM) in grapes. On the other hand, the team targeted DIPM-1, DIPM-2 and DIPM-4 in apple, in hopes of increasing resistance to fire blight disease. The targeted mutagenesis insertion and deletion (indel) rate was analyzed using targeted deep sequencing.

Their results show that direct delivery of CRISPR/Cas9 RNPs to the protoplast system enables targeted gene editing in grapevine and apple plants. This study is the first report of successful use of CRISPR/Cas9 RNPs-mediated protoplast transformation in grapevine and apple cultivars.

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