Precise genome modification in the crop species Zea mays using zinc-finger nucleases

Precise genome modification in the crop species Zea mays using zinc-finger nucleases Agricultural biotechnology is limited by the inefficiencies of conventional random mutagenesis and transgenesis. Because targeted genome modification in plants has been intractable , plant trait engineering remains a laborious, time-consuming and unpredictable undertaking. Here we report a broadly applicable, versatile solution to this problem: the use of designed zinc-finger nucleases (ZFNs) that induce a double-stranded break at their target locus . We describe the use of ZFNs to modify endogenous loci in plants of the crop species Zea mays . We show that simultaneous expression of ZFNs and delivery of a simple heterologous donor molecule leads to precise targeted addition of an herbicide-tolerance gene at the intended locus in a significant number of isolated events. ZFN-modified maize plants faithfully transmit these genetic changes to the next generation. Insertional disruption of one target locus, IPK1 , results in both herbicide tolerance and the expected alteration of the inositol phosphate profile in developing seeds. ZFNs can be used in any plant species amenable to DNA delivery; our results therefore establish a new strategy for plant genetic manipulation in basic science and agricultural applications. Current approaches to the challenge of improving agricultural productivity and global food production (for example, enhancing yield http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Nature Springer Journals

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Publisher
Springer Journals
Copyright
Copyright © 2009 Macmillan Publishers Limited. All rights reserved
ISSN
0028-0836
eISSN
1476-4687
DOI
10.1038/nature07992
Publisher site
See Article on Publisher Site

Abstract

Agricultural biotechnology is limited by the inefficiencies of conventional random mutagenesis and transgenesis. Because targeted genome modification in plants has been intractable , plant trait engineering remains a laborious, time-consuming and unpredictable undertaking. Here we report a broadly applicable, versatile solution to this problem: the use of designed zinc-finger nucleases (ZFNs) that induce a double-stranded break at their target locus . We describe the use of ZFNs to modify endogenous loci in plants of the crop species Zea mays . We show that simultaneous expression of ZFNs and delivery of a simple heterologous donor molecule leads to precise targeted addition of an herbicide-tolerance gene at the intended locus in a significant number of isolated events. ZFN-modified maize plants faithfully transmit these genetic changes to the next generation. Insertional disruption of one target locus, IPK1 , results in both herbicide tolerance and the expected alteration of the inositol phosphate profile in developing seeds. ZFNs can be used in any plant species amenable to DNA delivery; our results therefore establish a new strategy for plant genetic manipulation in basic science and agricultural applications. Current approaches to the challenge of improving agricultural productivity and global food production (for example, enhancing yield

Journal

NatureSpringer Journals

Published: Apr 29, 2009

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