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Related Experiment Video

Updated: May 24, 2025

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Transgene-free Genome Editing in Grapevine.

Edoardo Bertini1, Erica D'Incà1, Stefania Zattoni2

  • 1EdiVite s.r.l., Quartiere San Mauro 30, San Pietro Viminario, Padova, Italy.

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|March 3, 2025
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Summary

Researchers developed an efficient CRISPR/Cas9 genome editing protocol for grapevine (Vitis vinifera). This method enables DNA-free editing in protoplasts, allowing regeneration into whole plants for improved crop genetics.

Keywords:
CRISPR/Cas9DNA freeEmbryogenic callusGenome editingGrapevinePlant regenerationProtoplastsRibonucleoprotein complexes

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Area of Science:

  • Plant Biotechnology
  • Genome Editing
  • Crop Improvement

Background:

  • CRISPR/Cas9 technology offers precise genetic modification in plant breeding.
  • Previous grapevine editing attempts using ribonucleoprotein complexes lacked successful whole-plant regeneration.
  • Regenerating edited protoplasts into whole plants is crucial for practical application.

Purpose of the Study:

  • To establish an efficient protocol for DNA-free genome editing in grapevine (Vitis vinifera).
  • To achieve regeneration of CRISPR/Cas9-edited grapevine protoplasts into whole plants.
  • To provide a method for transgene/DNA-free genetic improvement in grapevine.

Main Methods:

  • Isolation of protoplasts from grapevine embryogenic callus.
  • Transfection of protoplasts with Cas9-single guide RNA (sgRNA) ribonucleoprotein (RNP) complexes.
  • Regeneration of edited protoplasts into whole grapevine plants.

Main Results:

  • Successful regeneration of whole grapevine plants from edited protoplasts.
  • Achieved transgene/DNA-free genome editing, avoiding exogenous DNA integration.
  • Regenerated plants exhibited normal morphology and growth habits, comparable to wild-type.

Conclusions:

  • This protocol enables highly efficient, DNA-free genome editing in grapevine.
  • The method addresses regulatory concerns by eliminating foreign DNA.
  • Facilitates genetic improvement of grapevine and other woody crops through precise editing.