Repositorium
Journal Article / 2009
Townsend, Jeffrey A.; Wright, David A.; Winfrey, Ronnie J.; Fu, Fengli; Maeder, Morgan L.; Joung, J. Keith; Voytas, Daniel F.
An efficient method for making directed DNA sequence modifications to plant genes (gene targeting) is at present lacking, thereby frustrating efforts to dissect plant gene function and engineer crop plants that better meet the world’s burgeoning need for food, fibre and fuel. Zinc-finger nucleases (ZFNs)—enzymes engineered to create DNA double-strand breaks at specific loci—are potent stimulators of gene targeting1,2 ; for example, they can be used to precisely modify engineered reporter genes in plants3,4 . Here we demonstrate high-frequency ZFN-stimulated gene targeting at endogenous plant genes, namely the tobacco acetolactate synthase genes (ALS SuRA and SuRB), for which specific mutations are known to confer resistance to imidazolinone and sulphonylurea herbicides5 . Herbicide-resistance mutations were introduced into SuR loci by ZFN-mediated gene targeting at frequencies exceeding 2% of transformed cells for mutations as far as 1.3 kilobases from the ZFN cleavage site. More than 40% of recombinant plants had modifications in multiple SuR alleles. The observed high frequency ofgene targeting indicates that it is nowpossible to efficientlymake targeted sequence changes in endogenous plant genes.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
499 |
Voytas, Daniel F. USA |
Tobacco |
Zinc-finger nucleases SuRA |
herbicide tolerance SDN1 |
Basic research Basic research |
500 |
Voytas, Daniel F. USA |
Tobacco |
Zinc-finger nucleases SuRB |
herbicide tolerance SDN1 |
Basic research Basic research |
501 |
Voytas, Daniel F. USA |
Tobacco |
Zinc-finger nucleases SuRA |
herbicide tolerance SDN2 |
Basic research Basic research |
502 |
Voytas, Daniel F. USA |
Tobacco |
Zinc-finger nucleases SuRB |
herbicide tolerance SDN2 |
Basic research Basic research |