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

被引：581

作者：

Shukla, Vipula K. ^{[1
]}

Doyon, Yannick ^{[2
]}

Miller, Jeffrey C. ^{[2
]}

DeKelver, Russell C. ^{[2
]}

Moehle, Erica A. ^{[2
]}

Worden, Sarah E. ^{[1
]}

Mitchell, Jon C. ^{[1
]}

Arnold, Nicole L. ^{[1
]}

Gopalan, Sunita ^{[2
]}

Meng, Xiangdong ^{[2
]}

Choi, Vivian M. ^{[2
]}

Rock, Jeremy M. ^{[2
]}

Wu, Ying-Ying ^{[2
]}

Katibah, George E. ^{[2
]}

Zhifang, Gao ^{[1
]}

McCaskill, David ^{[1
]}

Simpson, Matthew A. ^{[1
]}

Blakeslee, Beth ^{[1
]}

Greenwalt, Scott A. ^{[1
]}

Butler, Holly J. ^{[1
]}

Hinkley, Sarah J. ^{[2
]}

Zhang, Lei ^{[2
]}

Rebar, Edward J. ^{[2
]}

Gregory, Philip D. ^{[2
]}

Urnov, Fyodor D. ^{[2
]}

机构：

[1] Dow AgroSci, Indianapolis, IN 46268 USA

[2] Point Richmond Tech Ctr, Richmond, CA 94804 USA

来源：

NATURE | 2009年 / 459卷 / 7245期

关键词：

HOMOLOGOUS RECOMBINATION; MEDIATED TRANSFORMATION; GENE; EXPRESSION; MAIZE; CLEAVAGE; SEQUENCE; CELLS; MUTAGENESIS; DISRUPTION;

D O I：

10.1038/nature07992

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：437 / U156

页数：7

共 44 条

[1]

[Anonymous], 1989, Molecular Cloning: A Laboratory Manual

[2]

Armstrong C. L., 1991, Maize Genetics Cooperation News Letter, P92

[3] Stimulation of homologous recombination through targeted cleavage by chimeric nucleases [J].