Efficient Genome Editing in Apple Using a CRISPR/Cas9 system View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2016-08-17

AUTHORS

Chikako Nishitani, Narumi Hirai, Sadao Komori, Masato Wada, Kazuma Okada, Keishi Osakabe, Toshiya Yamamoto, Yuriko Osakabe

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. More... »

PAGES

31481

References to SciGraph publications

  • 2013-08-08. Targeted genome modification of crop plants using a CRISPR-Cas system in NATURE BIOTECHNOLOGY
  • 2013-08-08. Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease in NATURE BIOTECHNOLOGY
  • 2013-08-08. Multiplex and homologous recombination–mediated genome editing in Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9 in NATURE BIOTECHNOLOGY
  • 2009-03-15. Translocation of cell-penetrating peptides and delivery of their cargoes in triticale microspores in PLANT CELL REPORTS
  • 2003-10-15. Self-fertile apple resulting from S-RNase gene silencing in PLANT CELL REPORTS
  • 2007-05-08. Disruption of phytoene desaturase gene results in albino and dwarf phenotypes in Arabidopsis by impairing chlorophyll, carotenoid, and gibberellin biosynthesis in CELL RESEARCH
  • 2001. Incompatibility and Incongruity in Wild and Cultivated Plants in NONE
  • 2014-01-26. Improving CRISPR-Cas nuclease specificity using truncated guide RNAs in NATURE BIOTECHNOLOGY
  • 2015-10-19. DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins in NATURE BIOTECHNOLOGY
  • 2010-08-29. The genome of the domesticated apple (Malus × domestica Borkh.) in NATURE GENETICS
  • 1996-03. Self-incompatibility (S) alleles of the rosaceae encode members of a distinct class of the T2/S ribonuclease superfamily in MOLECULAR GENETICS AND GENOMICS
  • 2016-05-26. Optimization of CRISPR/Cas9 genome editing to modify abiotic stress responses in plants in SCIENTIFIC REPORTS
  • 2015-07-20. Efficient CRISPR/Cas9-mediated Targeted Mutagenesis in Populus in the First Generation in SCIENTIFIC REPORTS
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/srep31481

    DOI

    http://dx.doi.org/10.1038/srep31481

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1019790592

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/27530958


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