title: Improved specificity of TALE-based genome editing using an expanded RVD repertoire.
abstract: Transcription activator-like effector (TALE) proteins have gained broad appeal as a platform for targeted DNA recognition, largely owing to their simple rules for design. These rules relate the base specified by a single TALE repeat to the identity of two key residues (the repeat variable diresidue, or RVD) and enable design for new sequence targets via modular shuffling of these units. A key limitation of these rules is that their simplicity precludes options for improving designs that are insufficiently active or specific. Here we address this limitation by developing an expanded set of RVDs and applying them to improve the performance of previously described TALEs. As an extreme example, total conversion of a TALE nuclease to new RVDs substantially reduced off-target cleavage in cellular studies. By providing new RVDs and design strategies, these studies establish options for developing improved TALEs for broader application across medicine and biotechnology.
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Jeffrey C
authors: JC
authors: Miller
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Lei
authors: L
authors: Zhang
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Danny F
authors: DF
authors: Xia
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: John J
authors: JJ
authors: Campo
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Irina V
authors: IV
authors: Ankoudinova
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Dmitry Y
authors: DY
authors: Guschin
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Joshua E
authors: JE
authors: Babiarz
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Xiangdong
authors: X
authors: Meng
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Sarah J
authors: SJ
authors: Hinkley
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Stephen C
authors: SC
authors: Lam
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: David E
authors: DE
authors: Paschon
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Anna I
authors: AI
authors: Vincent
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Gladys P
authors: GP
authors: Dulay
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Kyle A
authors: KA
authors: Barlow
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: David A
authors: DA
authors: Shivak
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Elo
authors: E
authors: Leung
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Jinwon D
authors: JD
authors: Kim
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Rainier
authors: R
authors: Amora
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Fyodor D
authors: FD
authors: Urnov
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Philip D
authors: PD
authors: Gregory
authors: Sangamo BioSciences Inc., Richmond, California, USA.
authors: Edward J
authors: EJ
authors: Rebar
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation / Q000502:physiology*; D005819:Genetic Markers; D016678:Genome; D017393:RNA Editing / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D005819:Genetic Markers; D014157:Transcription Factors; D004247:DNA