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Sci Rep. 2016 Aug 17;6:31481. doi: 10.1038/srep31481.

Efficient Genome Editing in Apple Using a CRISPR/Cas9 system.

Author information

1
NARO Institute of Fruit Tree and Tea Science, 2-1 Fujimoto, Tsukuba, Ibaraki 305-8605, Japan.
2
Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, Iwate 020-8550, Japan.
3
NARO Institute of Fruit Tree and Tea Science, 92-24, Nabeyashiki, Shimokuriyagawa, Morioka, Iwate 020-0123, Japan.
4
Faculty of Bioscience and Bioindustry, Tokushima University, 2-1 Josanjima, Tokushima 770-8513, Japan.

Abstract

Genome editing is a powerful technique for genome modification in molecular research and crop breeding, and has the great advantage of imparting novel desired traits to genetic resources. However, the genome editing of fruit tree plantlets remains to be established. In this study, we describe induction of a targeted gene mutation in the endogenous apple phytoene desaturase (PDS) gene using the CRISPR/Cas9 system. Four guide RNAs (gRNAs) were designed and stably transformed with Cas9 separately in apple. Clear and partial albino phenotypes were observed in 31.8% of regenerated plantlets for one gRNA, and bi-allelic mutations in apple PDS were confirmed by DNA sequencing. In addition, an 18-bp gRNA also induced a targeted mutation. These CRIPSR/Cas9 induced-mutations in the apple genome suggest activation of the NHEJ pathway, but with some involvement also of the HR pathway. Our results demonstrate that genome editing can be practically applied to modify the apple genome.

PMID:
27530958
PMCID:
PMC4987624
DOI:
10.1038/srep31481
[Indexed for MEDLINE]
Free PMC Article

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