title: Structure based design of protein linkers for zinc finger nuclease.
abstract: Zinc finger nucleases are a promising tool to edit DNA in many biological applications, in particular for gene knockout. Despite many efforts the number of genes that can be effectively targeted with ZFNs remains severely limited, as available constructs cannot address arbitrary gene sequences. Here, we develop a novel concept to significantly enhance the number of DNA sequences that can be targeted by ZFN. Using an efficient computational model, we provide an extensive library of possible linker molecules between individual zinc finger motifs in the construct that can skip up to 10 base pairs between adjacent zinc finger recognition sites in the DNA sequence, which increases the number of genes that can be efficiently targeted by more than an order of magnitude.
authors.affiliation: Institute of Nanotechnology, Karlsruhe Institute of Technology, 76021 Karlsruhe, Germany.
authors.forename: Priya
authors.initials: P
authors.lastname: Anand
authors.forename: Alexander
authors.initials: A
authors.lastname: Schug
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wenzel
mesh_terms: D004720:Endonucleases / Q000737:chemistry*; D055786:Gene Knockout Techniques; D008958:Models, Molecular; D011487:Protein Conformation; D016335:Zinc Fingers
chemical_list: D004720:Endonucleases
keywords: CRISPR; Gene therapy; Krüppel type linker; Linker design; MOE; TALENs; ZFM; ZFN; ZFP; Zif268; Zinc finger nuclease; base pairs; bp; clustered regularly interspaced short palindromic repeats; molecular operating environment; transcription activator-like effector nucleases; zinc finger nucleases; zinc finger proteins; zinc-finger motifs