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.



[journal.frontiersin.org]
[journal.frontiersin.org]