Premium
Localized egg‐cell expression of effector proteins for targeted modification of the Arabidopsis genome
Author(s) -
EvenFaitelson Liron,
Samach Aviva,
MelamedBessudo Cathy,
AviviRagolsky Naomi,
Levy Avraham A.
Publication year - 2011
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2011.04741.x
Subject(s) - arabidopsis , biology , mutagenesis , gene , effector , computational biology , gene targeting , genome , genetics , homologous recombination , enhancer , genome engineering , genome editing , gene expression , microbiology and biotechnology , mutant
Summary 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.