caa a22 4500 606209263 CHVBK 20210128101027.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s00425-014-2224-x doi (NATIONALLICENCE)springer-10.1007/s00425-014-2224-x Targeted mutagenesis using zinc-finger nucleases in perennial fruit trees [Elektronische Daten] [Reut Peer, Gil Rivlin, Sara Golobovitch, Moshe Lapidot, Amit Gal-On, Alexander Vainstein, Tzvi Tzfira, Moshe Flaishman] Main conclusion : Targeting a gene in apple or fig with ZFN, introduced by transient or stable transformation, should allow genome editing with high precision to advance basic science and breeding programs. Genome editing is a powerful tool for precise gene manipulation in any organism; it has recently been shown to be of great value for annual plants. Classical breeding strategies using conventional cross-breeding and induced mutations have played an important role in the development of new cultivars in fruit trees. However, fruit-tree breeding is a lengthy process with many limitations. Efficient and widely applied methods for targeted modification of fruit-tree genomes are not yet available. In this study, transgenic apple and fig lines carrying a zinc-finger nuclease (ZFNs) under the control of a heat-shock promoter were developed. Editing of a mutated uidA gene, following expression of the ZFN genes by heat shock, was confirmed by GUS staining and PCR product sequencing. Finally, whole plants with a repaired uidA gene due to deletion of a stop codon were regenerated. The ZFN-mediated gene modifications were stable and passed onto regenerants from ZFN-treated tissue cultures. This is the first demonstration of efficient and precise genome editing, using ZFN at a specific genomic locus, in two different perennial fruit trees—apple and fig. We conclude that targeting a gene in apple or fig with a ZFN introduced by transient or stable transformation should allow knockout of a gene of interest. Using this technology for genome editing allows for marker gene-independent and antibiotic selection-free genome engineering with high precision in fruit trees to advance basic science as well as nontransgenic breeding programs. Springer-Verlag Berlin Heidelberg, 2014 Targeted genome editing nationallicence Zinc-finger nuclease (ZFN) nationallicence Perennial fruit trees nationallicence Apple ( Malus domestica ) nationallicence Fig ( Ficus carica ) nationallicence Peer Reut Institute of Plant Sciences, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut Rivlin Gil Institute of Plant Sciences, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut Golobovitch Sara Institute of Plant Sciences, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut Lapidot Moshe Institute of Plant Sciences, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut Gal-On Amit Department of Plant Pathology and Weed Science, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut Vainstein Alexander Institute of Plant Sciences and Genetics in Agriculture, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel aut Tzfira Tzvi Department of Life Sciences, Ben-Gurion University of the Negev, 84105, Beersheba, Israel aut Flaishman Moshe Institute of Plant Sciences, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut Planta Springer Berlin Heidelberg 241/4(2015-04-01), 941-951 0032-0935 241:4<941 2015 241 425 https://doi.org/10.1007/s00425-014-2224-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00425-014-2224-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Peer Reut Institute of Plant Sciences, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut NATIONALLICENCE 700 1- Rivlin Gil Institute of Plant Sciences, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut NATIONALLICENCE 700 1- Golobovitch Sara Institute of Plant Sciences, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut NATIONALLICENCE 700 1- Lapidot Moshe Institute of Plant Sciences, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut NATIONALLICENCE 700 1- Gal-On Amit Department of Plant Pathology and Weed Science, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut NATIONALLICENCE 700 1- Vainstein Alexander Institute of Plant Sciences and Genetics in Agriculture, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel aut NATIONALLICENCE 700 1- Tzfira Tzvi Department of Life Sciences, Ben-Gurion University of the Negev, 84105, Beersheba, Israel aut NATIONALLICENCE 700 1- Flaishman Moshe Institute of Plant Sciences, Agricultural Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel aut NATIONALLICENCE 773 0- Planta Springer Berlin Heidelberg 241/4(2015-04-01), 941-951 0032-0935 241:4<941 2015 241 425