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The most common applications for engineered zinc finger arrays include zinc finger transcription factors and zinc finger nucleases, but other applications have also been described. Typical engineered zinc finger arrays have between 3 and 6 individual zinc finger motifs and bind target sites ranging from 9 basepairs to 18 basepairs in length.
Zinc finger nucleases have also been used in a mouse model of haemophilia [31] and a clinical trial found CD4+ human T-cells with the CCR5 gene disrupted by zinc finger nucleases to be safe as a potential treatment for HIV/AIDS. [32] ZFNs are also used to create a new generation of genetic disease models called isogenic human disease models.
Zinc finger nucleases are genetically engineered enzymes that combine fusing a zinc finger DNA-binding domain on a DNA-cleavage domain. These are also combined with CRISPR-CAS9 or TALENs to gain a sequence-specific addition, or deletion, within the genome of more complex cells and organisms. [17]
In the early 2000s, German researchers began developing zinc finger nucleases (ZFNs), synthetic proteins whose DNA-binding domains enable them to create double-stranded breaks in DNA at specific points. ZFNs have a higher precision and the advantage of being smaller than Cas9, but ZFNs are not as commonly used as CRISPR-based methods.
TALENs are less efficient than CRISPR/Cas9, but they are still a useful tool for genome editing. Zinc finger editing, using zinc finger nucleases Natural genetic engineering (NGE) has been proposed by molecular biologist James A. Shapiro to account for novelty created in the course of biological evolution.
A subsequent study used modular assembly to generate zinc finger nucleases with both 3-finger arrays and 4-finger arrays and observed a much higher success rate with 4-finger arrays. [29] A variant of modular assembly that takes the context of neighboring fingers into account has also been reported and this method tends to yield proteins with ...
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