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Related Experiment Video

Updated: May 3, 2026

Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector

Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector

Published on: March 28, 2018

DNA replicons for plant genome engineering.

Nicholas J Baltes¹, Javier Gil-Humanes, Tomas Cermak

¹Department of Genetics, Cell Biology, and Development, Center for Genome Engineering, University of Minesota, Mineapolis, Minesota 55455.

|January 21, 2014

Summary

This summary is machine-generated.

Geminivirus replicons significantly enhance plant genome engineering by enabling efficient delivery of gene editing tools and DNA repair templates. This method accelerates the generation of genetically modified plants with desired DNA sequence modifications.

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Area of Science:

Plant biotechnology
Molecular biology
Genomics

Background:

Targeted modification of plant genomes is hindered by inefficient delivery methods for genome engineering reagents.
Sequence-specific nucleases (e.g., ZFNs, TALENs, CRISPR/Cas) are powerful tools for DNA editing.

Purpose of the Study:

To develop and validate geminivirus-based replicons for transient expression of nucleases and delivery of repair templates in plants.
To improve the efficiency and speed of plant genome engineering.

Main Methods:

Utilized geminivirus-based replicons for transient expression of zinc-finger nucleases, transcription activator-like effector nucleases, and CRISPR/Cas systems.
Delivered DNA repair templates alongside nucleases using the geminivirus replicon system.
Employed a general plant transformation method for regeneration of modified plantlets.

Main Results:

Geminivirus replicons enhanced gene targeting frequencies in tobacco (Nicotiana tabacum) by one to two orders of magnitude compared to Agrobacterium tumefaciens T-DNA.
Replication of the repair template and geminivirus replication initiator proteins promoted gene targeting.
Generated plants with desired DNA sequence modifications in under 6 weeks.

Conclusions:

Geminivirus replicons offer a highly efficient and versatile platform for plant genome engineering.
This approach overcomes previous limitations in delivering genome editing reagents to plant cells.
The broad host range of geminiviruses suggests wide applicability for this method in plant biotechnology.