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.
Shukla
Vipula K
VK
Dow AgroSciences, 9330 Zionsville Road, Indianapolis, Indiana 46268, USA. vkshukla@dow.com
Doyon
Yannick
Y
Miller
Jeffrey C
JC
DeKelver
Russell C
RC
Moehle
Erica A
EA
Worden
Sarah E
SE
Mitchell
Jon C
JC
Arnold
Nicole L
NL
Gopalan
Sunita
S
Meng
Xiangdong
X
Choi
Vivian M
VM
Rock
Jeremy M
JM
Wu
Ying-Ying
YY
Katibah
George E
GE
Zhifang
Gao
G
McCaskill
David
D
Simpson
Matthew A
MA
Blakeslee
Beth
B
Greenwalt
Scott A
SA
Butler
Holly J
HJ
Hinkley
Sarah J
SJ
Zhang
Lei
L
Rebar
Edward J
EJ
Gregory
Philip D
PD
Urnov
Fyodor D
FD
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
D006540:Herbicides; D007295:Inositol Phosphates; D003851:Deoxyribonucleases