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.
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