Activity and specificity of TRV-mediated gene editing in plants

Zahir Ali, Aala Abul-Faraj, Marek Piatek, Magdy M. Mahfouz*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

64 Scopus citations

Abstract

Plant trait engineering requires efficient targeted genome-editing technologies. Clustered regularly interspaced palindromic repeats (CRISPRs)/ CRISPR associated (Cas) type II system is used for targeted genome-editing applications across eukaryotic species including plants. Delivery of genome engineering reagents and recovery of mutants remain challenging tasks for in planta applications. Recently, we reported the development of Tobacco rattle virus (TRV)-mediated genome editing in Nicotiana benthamiana. TRV infects the growing points and possesses small genome size; which facilitate cloning, multiplexing, and agroinfections. Here, we report on the persistent activity and specificity of the TRV-mediated CRISPR/Cas9 system for targeted modification of the Nicotiana benthamiana genome. Our data reveal the persistence of the TRVmediated Cas9 activity for up to 30 d post-agroinefection. Further, our data indicate that TRV-mediated genome editing exhibited no off-target activities at potential off-targets indicating the precision of the system for plant genome engineering. Taken together, our data establish the feasibility and exciting possibilities of using virus-mediated CRISPR/Cas9 for targeted engineering of plant genomes.

Original languageEnglish (US)
Article numbere1044191
JournalPlant Signaling and Behavior
Volume10
Issue number10
DOIs
StatePublished - 2015

Bibliographical note

Publisher Copyright:
© 2015 Taylor and Francis Group, LLC.

Keywords

  • CRISPR/Cas9 system
  • Plant genome engineering
  • Synthetic site-specific nucleases (SSNs)
  • TRV
  • Viral-mediated genome editing

ASJC Scopus subject areas

  • Plant Science

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