A rapid, generally applicable method to engineer zinc fingers illustrated by targeting the HIV-1 promoter

@article{Isalan2001ARG,
  title={A rapid, generally applicable method to engineer zinc fingers illustrated by targeting the HIV-1 promoter},
  author={Mark Isalan and Aaron Klug and Yeng Cheng Choo},
  journal={Nature Biotechnology},
  year={2001},
  volume={19},
  pages={656-660}
}
DNA-binding domains with predetermined sequence specificity are engineered by selection of zinc finger modules using phage display, allowing the construction of customized transcription factors. Despite remarkable progress in this field, the available protein-engineering methods are deficient in many respects, thus hampering the applicability of the technique. Here we present a rapid and convenient method that can be used to design zinc finger proteins against a variety of DNA-binding sites… 
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The development of a specificity profiling system that facilitates rapid and inexpensive characterization of engineered zinc-finger modules and it is demonstrated that specificity data collected using this system can be employed to rationally design zinc fingers with improved DNA-binding specificities.
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  • 2010
TLDR
A bipartite phage display approach that enables selection and recombination of variants of zinc finger DNA-binding domains from a pair of premade complementary phage libraries for any given 9-bp DNA sequence is devised.
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TLDR
This chapter will describe methods and give guidance for the creation of ZFPs using MA, and describe a single-strand annealing recombination assay for the initial testing of zinc finger nucleases.
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TLDR
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  • 2010
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The zinc finger design can be used to construct DNA-binding proteins for specific intervention in gene expression, and several applications of such engineered ZFPs are described, including some of therapeutic importance, and also their adaptation for breeding improved crop plants.
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TLDR
This protocol describes the design and construction of new DNA-binding domains comprised of zinc fingers directed at selected DNA sequences, developed as targetable cleavage reagents for double-stranded DNA, both in vitro and in vivo.
A Phage Display System to Profile the DNA-binding Specificities of C2H2 Zinc Fingers
TLDR
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The discovery of zinc fingers and their applications in gene regulation and genome manipulation.
  • A. Klug
  • Biology, Chemistry
    Annual review of biochemistry
  • 2010
TLDR
The zinc finger design is ideally suited for engineering proteins to target specific genes, and several applications of such engineered zinc finger proteins are described here, including some of therapeutic importance.
DNA-binding specificity is a major determinant of the activity and toxicity of zinc-finger nucleases.
TLDR
The results of these cell-based assays reveal that the DNA-binding specificity--in addition to the affinity--is a major determinant of ZFN activity and is inversely correlated with ZFN-associated toxicity, and provide the first evidence that engineering strategies, which account for context-dependent DNA- binding effects, yield ZFs that function as highly efficient ZFNs in human cells.
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