title: Precise genome modification in the crop species Zea mays using zinc-finger nucleases.
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.
authors.affiliation: Dow AgroSciences, 9330 Zionsville Road, Indianapolis, Indiana 46268, USA. vkshukla@dow.com
authors.forename: Vipula K
authors.initials: VK
authors.lastname: Shukla
authors.forename: Yannick
authors.initials: Y
authors.lastname: Doyon
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Miller
authors.forename: Russell C
authors.initials: RC
authors.lastname: DeKelver
authors.forename: Erica A
authors.initials: EA
authors.lastname: Moehle
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Worden
authors.forename: Jon C
authors.initials: JC
authors.lastname: Mitchell
authors.forename: Nicole L
authors.initials: NL
authors.lastname: Arnold
authors.forename: Sunita
authors.initials: S
authors.lastname: Gopalan
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Meng
authors.forename: Vivian M
authors.initials: VM
authors.lastname: Choi
authors.forename: Jeremy M
authors.initials: JM
authors.lastname: Rock
authors.forename: Ying-Ying
authors.initials: YY
authors.lastname: Wu
authors.forename: George E
authors.initials: GE
authors.lastname: Katibah
authors.forename: Gao
authors.initials: G
authors.lastname: Zhifang
authors.forename: David
authors.initials: D
authors.lastname: McCaskill
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Simpson
authors.forename: Beth
authors.initials: B
authors.lastname: Blakeslee
authors.forename: Scott A
authors.initials: SA
authors.lastname: Greenwalt
authors.forename: Holly J
authors.initials: HJ
authors.lastname: Butler
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Hinkley
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Rebar
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.forename: Fyodor D
authors.initials: FD
authors.lastname: Urnov
mesh_terms: D001709:Biotechnology / Q000379:methods*; D003851:Deoxyribonucleases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D030841:Food, Genetically Modified; D018390:Gene Targeting / Q000379:methods*; D017343:Genes, Plant / Q000235:genetics; D018745:Genome, Plant / Q000235:genetics*; D053209:Herbicide Resistance / Q000235:genetics; D006540:Herbicides / Q000494:pharmacology; D040941:Heredity; D007295:Inositol Phosphates / Q000378:metabolism; D016297:Mutagenesis, Site-Directed / Q000379:methods; D030821:Plants, Genetically Modified; D011995:Recombination, Genetic / Q000235:genetics; D015203:Reproducibility of Results; D003313:Zea mays / Q000235:genetics*; D016335:Zinc Fingers
chemical_list: D006540:Herbicides; D007295:Inositol Phosphates; D003851:Deoxyribonucleases