title: Selection-free zinc-finger-nuclease engineering by context-dependent assembly (CoDA).
abstract: Engineered zinc-finger nucleases (ZFNs) enable targeted genome modification. Here we describe context-dependent assembly (CoDA), a platform for engineering ZFNs using only standard cloning techniques or custom DNA synthesis. Using CoDA-generated ZFNs, we rapidly altered 20 genes in Danio rerio, Arabidopsis thaliana and Glycine max. The simplicity and efficacy of CoDA will enable broad adoption of ZFN technology and make possible large-scale projects focused on multigene pathways or genome-wide alterations.
authors.affiliation: Molecular Pathology Unit and Center for Cancer Research, Massachusetts General Hospital, Charlestown, Massachusetts, USA.
authors.forename: Jeffry D
authors.initials: JD
authors.lastname: Sander
authors.forename: Elizabeth J
authors.initials: EJ
authors.lastname: Dahlborg
authors.forename: Mathew J
authors.initials: MJ
authors.lastname: Goodwin
authors.forename: Lindsay
authors.initials: L
authors.lastname: Cade
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.forename: Daniel
authors.initials: D
authors.lastname: Cifuentes
authors.forename: Shaun J
authors.initials: SJ
authors.lastname: Curtin
authors.forename: Jessica S
authors.initials: JS
authors.lastname: Blackburn
authors.forename: Stacey
authors.initials: S
authors.lastname: Thibodeau-Beganny
authors.forename: Yiping
authors.initials: Y
authors.lastname: Qi
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Pierick
authors.forename: Ellen
authors.initials: E
authors.lastname: Hoffman
authors.forename: Morgan L
authors.initials: ML
authors.lastname: Maeder
authors.forename: Cyd
authors.initials: C
authors.lastname: Khayter
authors.forename: Deepak
authors.initials: D
authors.lastname: Reyon
authors.forename: Drena
authors.initials: D
authors.lastname: Dobbs
authors.forename: David M
authors.initials: DM
authors.lastname: Langenau
authors.forename: Robert M
authors.initials: RM
authors.lastname: Stupar
authors.forename: Antonio J
authors.initials: AJ
authors.lastname: Giraldez
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Voytas
authors.forename: Randall T
authors.initials: RT
authors.lastname: Peterson
authors.forename: Jing-Ruey J
authors.initials: JR
authors.lastname: Yeh
authors.forename: J Keith
authors.initials: JK
authors.lastname: Joung
mesh_terms: D000818:Animals; D017360:Arabidopsis / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004720:Endonucleases / Q000235:genetics* / Q000378:metabolism*; D016678:Genome; D015202:Protein Engineering; D013025:Glycine max / Q000235:genetics; D015027:Zebrafish / Q000235:genetics; D016335:Zinc Fingers / Q000235:genetics / Q000502:physiology*
chemical_list: D004268:DNA-Binding Proteins; D004720:Endonucleases