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
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DOI：
10.1038/nature07992
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发表时间：
2009-05-21
期刊：
影响因子：
64.8
通讯作者：
Urnov, Fyodor D.
中科院分区：
文献类型：
--
作者：
Shukla, Vipula K.;Doyon, Yannick;Urnov, Fyodor D.
农业生物技术受到传统随机诱变和转基因技术效率低下的限制。由于植物中的靶向基因组修饰一直难以实现，植物性状工程仍然是一项费力、耗时且不可预测的工作。在此我们报道了一个对该问题广泛适用且通用的解决方案：使用设计的锌指核酸酶（ZFNs），其能在靶位点诱导双链断裂。我们描述了使用ZFNs来修饰作物玉米植物中的内源基因座。我们表明，ZFNs的同时表达以及一个简单的异源供体分子的导入，在大量独立事件中能在预期位点精确地靶向添加一个抗除草剂基因。经ZFN修饰的玉米植株能将这些遗传变化忠实地传递给下一代。一个靶位点IPK1的插入性破坏，导致了抗除草剂特性以及发育中种子的肌醇磷酸谱的预期改变。ZFNs可用于任何可进行DNA导入的植物物种；因此我们的结果为基础科学和农业应用中的植物基因操作建立了一种新策略。
Agricultural biotechnology is limited by the inefficiencies of conventional random mutagenesis and transgenesis. Because targeted genome modification in plants has been intractable(1), 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(2). 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.