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Virus-Induced Heritable Gene Editing in Plants.

Ugrappa Nagalakshmi1, Nathan Meier2, Savithramma P Dinesh-Kumar3

  • 1Department of Plant Biology and The Genome Center, College of Biological Sciences, University of California, Davis, Davis, CA, USA. unagalakshmi@ucdavis.edu.

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Summary

This study presents an optimized TRV-based viral vector for delivering CRISPR/Cas gene editing tools into plants. This method enables efficient, heritable gene editing in model plants like Nicotiana benthamiana and Arabidopsis thaliana.

Keywords:
ArabidopsisCas9Gene editingGuide RNAHeritableNicotiana benthamianaPhytoene desaturase (PDS)Tobacco Rattle Virus (TRV)

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Area of Science:

  • Plant biotechnology
  • Molecular biology
  • Genetics

Background:

  • CRISPR/Cas gene editing is crucial for plant gene function studies.
  • Delivering CRISPR/Cas components into plant cells remains a significant challenge, often requiring complex tissue culture methods.
  • Plant viral vectors offer a promising alternative for gene editing component delivery.

Purpose of the Study:

  • To develop an optimized protocol for efficient heritable gene editing in plants.
  • To overcome the limitations of traditional CRISPR/Cas delivery methods.
  • To establish a versatile viral vector system for studying gene function in model plants.

Main Methods:

  • Utilized a tobacco rattle virus (TRV)-based vector system.
  • Engineered the TRV vector to deliver single-guide RNA (sgRNA) sequences fused to mobile tRNA.
  • Applied the protocol to Nicotiana benthamiana and Arabidopsis thaliana.

Main Results:

  • Demonstrated efficient heritable gene editing in both Nicotiana benthamiana and Arabidopsis thaliana.
  • The TRV-based vector system successfully delivered sgRNAs for targeted gene modification.
  • The protocol provides a streamlined approach compared to traditional methods.

Conclusions:

  • The optimized TRV-based viral vector system facilitates efficient and heritable CRISPR/Cas gene editing in plants.
  • This method overcomes key bottlenecks in plant gene editing, simplifying the study of gene function.
  • The protocol is adaptable for investigating any gene of interest in various plant species.