Enhancing zinc-finger-nuclease activity with improved obligate heterodimeric architectures

@article{Doyon2011EnhancingZA,
  title={Enhancing zinc-finger-nuclease activity with improved obligate heterodimeric architectures},
  author={Yannick Doyon and Thuy D. Vo and Matthew C. Mendel and Shon G Greenberg and Jianbin Wang and Danny F Xia and Jeffrey C. Miller and Fyodor D. Urnov and Philip D. Gregory and Michael C. Holmes},
  journal={Nature Methods},
  year={2011},
  volume={8},
  pages={74-79}
}
Zinc-finger nucleases (ZFNs) drive efficient genome editing by introducing a double-strand break into the targeted gene. Cleavage is induced when two custom-designed ZFNs heterodimerize upon binding DNA to form a catalytically active nuclease complex. The importance of this dimerization event for subsequent cleavage activity has stimulated efforts to engineer the nuclease interface to prevent undesired homodimerization. Here we report the development and application of a yeast-based selection… 

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The mechanism of how it works, currently available target assessment, ZFP library construction and screening methods, target modification strategies, as well as a collection of specie and genes that have been successfully modified by ZFN are focused on.

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An expanded set of zinc finger nuclease architectures are developed that increase the available configurations by a factor of 64 and can target almost every base at loci of therapeutic significance and show that these new architectures may be used for targeting three loci with a high degree of precision, efficiency, and specificity.

Synthetic zinc finger nuclease design and rapid assembly.

This is the first demonstration of in silico-designed, oligonucleotide-assembled, synthetic ZFNs, requiring no specialized templates or reagents that are capable of endogenous human gene target site activity, and should facilitate a more widespread use of Z FNs in the research community.

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An approach for improving the specificity of zinc finger nucleases (ZFNs) that engineers the FokI catalytic domain with the aim of slowing cleavage, which should selectively reduce activity at low-affinity off-target sites is developed.

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