TY - JOUR
T1 - Localized egg-cell expression of effector proteins for targeted modification of the Arabidopsis genome
AU - Even-Faitelson, Liron
AU - Samach, Aviva
AU - Melamed-Bessudo, Cathy
AU - Avivi-Ragolsky, Naomi
AU - Levy, Avraham
N1 - BARD; EUWe would like to thank Ueli Grossniklauss and Nir Ohad for providing us with useful advice on egg cell-specific expression, and Tzvi Tzfira for fruitful discussions and his generosity with reagents. We thank Vladimir Kiss for help with the confocal microscopy, Michal Kenan-Eichler for construction of the new RFP GT cassette, and Michal Lieberman-Lazarovitch and members of the Levy lab for useful discussions. This work was supported by grants from BARD and by the EU-FP6 TAGIP and EU-FP7 RECBREED projects.
PY - 2011/12
Y1 - 2011/12
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=82355181082&partnerID=8YFLogxK
U2 - https://doi.org/10.1111/j.1365-313X.2011.04741.x
DO - https://doi.org/10.1111/j.1365-313X.2011.04741.x
M3 - مقالة
SN - 0960-7412
VL - 68
SP - 929
EP - 937
JO - Plant Journal
JF - Plant Journal
IS - 5
ER -