title: A TALE nuclease architecture for efficient genome editing.
abstract: Nucleases that cleave unique genomic sequences in living cells can be used for targeted gene editing and mutagenesis. Here we develop a strategy for generating such reagents based on transcription activator-like effector (TALE) proteins from Xanthomonas. We identify TALE truncation variants that efficiently cleave DNA when linked to the catalytic domain of FokI and use these nucleases to generate discrete edits or small deletions within endogenous human NTF3 and CCR5 genes at efficiencies of up to 25%. We further show that designed TALEs can regulate endogenous mammalian genes. These studies demonstrate the effective application of designed TALE transcription factors and nucleases for the targeted regulation and modification of endogenous genes.
authors.affiliation: Sangamo BioSciences, Inc., Richmond, California, USA.
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Miller
authors.forename: Siyuan
authors.initials: S
authors.lastname: Tan
authors.forename: Guijuan
authors.initials: G
authors.lastname: Qiao
authors.forename: Kyle A
authors.initials: KA
authors.lastname: Barlow
authors.forename: Jianbin
authors.initials: J
authors.lastname: Wang
authors.forename: Danny F
authors.initials: DF
authors.lastname: Xia
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Meng
authors.forename: David E
authors.initials: DE
authors.lastname: Paschon
authors.forename: Elo
authors.initials: E
authors.lastname: Leung
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Hinkley
authors.forename: Gladys P
authors.initials: GP
authors.lastname: Dulay
authors.forename: Kevin L
authors.initials: KL
authors.lastname: Hua
authors.forename: Irina
authors.initials: I
authors.lastname: Ankoudinova
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Cost
authors.forename: Fyodor D
authors.initials: FD
authors.lastname: Urnov
authors.forename: H Steve
authors.initials: HS
authors.lastname: Zhang
authors.forename: Michael C
authors.initials: MC
authors.lastname: Holmes
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Rebar
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D004247:DNA / Q000235:genetics / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics / Q000378:metabolism; D005818:Genetic Engineering; D016678:Genome; D006801:Humans; D020014:K562 Cells; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D019713:Receptors, CCR5 / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics; D014974:Xanthomonas
chemical_list: D001426:Bacterial Proteins; D019713:Receptors, CCR5; D014157:Transcription Factors; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D004247:DNA; C035042:endodeoxyribonuclease FokI; D015252:Deoxyribonucleases, Type II Site-Specific