Electronic Design and Structure Simulation of Artificial Zinc Finger Nucleases

Taking the electronic design of zinc finger nucleases(ZFN) for the multiple locus gene targeting of pig as an example,the electronic design of an ideal optimized artificial ZFN was explored to reduce the toxicity of the ZFN.The method of the OPEN electronic design was used.The three-dimensional structure of zinc finger protein(ZFP) was predicted by the methods of bioinformatics.The appropriate linkers and spacers were designed and optimized.The heterodimer of Fok I was mutated and optimized.Three-finger zinc finger sequences(ZF) were designed by OPEN method and the ZFs were connected to ZFP sequence.The AT-rich spacers with 6 bp were designed.The inter-domain linkers with four amino acid residues were connected to ZFP.The cutting domain of Fok Ⅰwith double mutant heterodimers were connected to the ZFP too.The spatial structure of ZFN was simulated and predicted by a variety of software tools.The efficient of the designed ZFN was more than 50%.The programs of electronic design of ideal ZFN including ZFP specifically bind to DNA,the linkers and spacers with the optimal length and combination,and the optimal Fok I heterodimer domain with specific cutting were explored and obtained.