These authors contributed equally to this work.
Localized egg-cell expression of effector proteins for targeted modification of the Arabidopsis genome
Article first published online: 4 OCT 2011
© 2011 The Authors. The Plant Journal © 2011 Blackwell Publishing Ltd
The Plant Journal
Volume 68, Issue 5, pages 929–937, December 2011
How to Cite
Even-Faitelson, L., Samach, A., Melamed-Bessudo, C., Avivi-Ragolsky, N. and Levy, A. A. (2011), Localized egg-cell expression of effector proteins for targeted modification of the Arabidopsis genome. The Plant Journal, 68: 929–937. doi: 10.1111/j.1365-313X.2011.04741.x
- Issue published online: 25 NOV 2011
- Article first published online: 4 OCT 2011
- Accepted manuscript online: 16 AUG 2011 09:14AM EST
- Received 20 June 2011; revised 10 August 2011; accepted 11 August 2011; published online 4 October 2011.
- targeted mutagenesis;
- gene targeting;
- egg-cell enhancer;
- zinc-finger nuclease;
Targeted modification of the genome is an important genetic tool, which can be achieved via homologous, non-homologous or site-specific recombination. Although numerous efforts have been made, such a tool does not exist for routine applications in plants. This work describes a simple and useful method for targeted mutagenesis or gene targeting, tailored to floral-dip transformation in Arabidopsis, by means of specific protein expression in the egg cell. Proteins stably or transiently expressed under the egg apparatus-specific enhancer (EASE) were successfully localized to the area of the egg cell. Moreover, a zinc-finger nuclease expressed under EASE induced targeted mutagenesis. Mutations obtained under EASE control corresponded to genetically independent events that took place specifically in the germline. In addition, RAD54 expression under EASE led to an approximately 10-fold increase in gene targeting efficiency, when compared with wild-type plants. EASE-controlled gene expression provides a method for the precise engineering of the Arabidopsis genome through temporally and spatially controlled protein expression. This system can be implemented as a useful method for basic research in Arabidopsis, as well as in the optimization of tools for targeted genetic modifications in crop plants.