DNA Replicons for Plant Genome Engineering  . Issue 1 (17th January 2014)

• Record Type: Journal Article
• Authors: Baltes, Nicholas J.; Gil-Humanes, Javier; Cermak, Tomas; Atkins, Paul A.; Voytas, Daniel F.
• Is Part Of: The Plant Cell. Volume 26:Issue 1(2014)
• Page Start: 151
• Page End: 163
• Publication Date: 2014-01-17
• DOI: 10.1105/tpc.113.119792
• Languages: English
• ISSNs: 1040-4651

Abstract

Abstract : The ability to precisely modify DNA in cells offers great opportunities for basic and applied research, yet it remains difficult to achieve for most plant species. This work explores the use of geminiviruses for genome engineering. Properties of geminiviruses, including extrachromosomal replication and pleiotropic activity of virus proteins, enabled highly efficient editing of the tobacco genome. Abstract: Sequence-specific nucleases enable facile editing of higher eukaryotic genomic DNA; however, targeted modification of plant genomes remains challenging due to ineffective methods for delivering reagents for genome engineering to plant cells. Here, we use geminivirus-based replicons for transient expression of sequence-specific nucleases (zinc-finger nucleases, transcription activator–like effector nucleases, and the clustered, regularly interspaced, short palindromic repeat/Cas system) and delivery of DNA repair templates. In tobacco ( Nicotiana tabacum ), replicons based on the bean yellow dwarf virus enhanced gene targeting frequencies one to two orders of magnitude over conventional Agrobacterium tumefaciens T-DNA. In addition to the nuclease-mediated DNA double-strand breaks, gene targeting was promoted by replication of the repair template and pleiotropic activity of the geminivirus replication initiator proteins. We demonstrate the feasibility of using geminivirus replicons to generate plants with a desired DNA sequence modification. By adopting a general plant transformation method, plantlets with a desired DNA change were regenerated in <6 weeks. These results, in addition to the large host range of geminiviruses, advocate the use of replicons for plant genome engineering.