Sage-Tips

How can zinc finger domains be used to develop gene editing technologies?

Zinc finger domains can be engineered to target specific desired DNA sequences and this enables zinc-finger nucleases to target unique sequences within complex genomes. By taking advantage of endogenous DNA repair machinery, these reagents can be used to precisely alter the genomes of higher organisms.

How does the zinc finger actually bind to the DNA?

The binding of zinc finger is found to be distinct from many other DNA-binding proteins that bind DNA through the 2-fold symmetry of the double helix, instead zinc fingers are linked linearly in tandem to bind nucleic acid sequences of varying lengths. Zinc fingers often bind to a sequence of DNA known as the GC box.

How does a zinc atom help stabilize the zinc finger?

The zinc atom is simultaneously bound by the 2 cysteine and the 2 histidine side chains. DNA has a negatively-charged phosphate backbone. Therefore, the positively- charged arginine of the zinc finger can bind to DNA via an electrostatic interaction.

Which protein contains a zinc finger domain?

Zinc finger domains are one of the most common structural motifs in eukaryotic cells, which employ the motif in some of their most important proteins (including TFIIIA, CTCF, and ZiF268). These DNA binding proteins contain up to 37 zinc finger domains connected by flexible linker regions.

What is zinc finger gene editing?

Zinc finger nucleases (ZFNs) are a class of engineered DNA-binding proteins that facilitate targeted editing of the genome by creating double-strand breaks in DNA at user-specified locations.

How does zinc finger nucleases recognize specific DNA sequences?

ZFNs use DNA binding domains to recognize ~ 3 bp sequences that are joined together to generate arrays which allows to target desired DNA sequences. TALENs bind to DNA using TAL effector repeats that are ligated together and generate arrays that allows to recognize target DNA sequences.

What does zinc bind with?

Liganding residues. The binding residues that dominate zinc binding sites are well established: cysteine, histidine and the acidic residues aspartate and glutamate (7).

What is the chemical basis of zinc binding by a zinc finger?

Zinc finger protein (ZFP) is a common DNA binding domain found in many transcription factors. It consists of ∼30 amino acids that may recognize three base pairs of DNA. Combining 3–6 ZFP in a molecule, it can recognize and bind to a specific DNA into the genome.

What binds to zinc?

Common Zn2+ ligands found within proteins include cysteine (S), histidine (N), aspartate (O), and glutamate (O) residues. The ionization state of the thiol group of cysteine governs its ability to bind metals, including Zn2+.

What is the function of zinc finger nucleases?

What is the purpose of genome editing?

Genome editing, also called gene editing, is an area of research seeking to modify genes of living organisms to improve our understanding of gene function and develop ways to use it to treat genetic or acquired diseases.

What is a DNA cleavage domain?

A reaction that severs one of the covalent sugar-phosphate linkages between NUCLEOTIDES that compose the sugar phosphate backbone of DNA. It is catalyzed enzymatically, chemically or by radiation. Cleavage may be exonucleolytic – removing the end nucleotide, or endonucleolytic – splitting the strand in two.

What are zinc- finger nucleases (ZFNs)?

Jump to navigation Jump to search. Zinc-finger nucleases (ZFNs) are artificial restriction enzymes generated by fusing a zinc finger DNA-binding domain to a DNA-cleavage domain. Zinc finger domains can be engineered to target specific desired DNA sequences and this enables zinc-finger nucleases to target unique sequences within complex genomes.

How are zinc-fingers made?

The most straightforward method to generate new zinc-finger arrays is to combine smaller zinc-finger “modules” of known specificity. The most common modular assembly process involves combining three separate zinc fingers that can each recognize a 3 basepair DNA sequence to generate a 3-finger array that can recognize a 9 basepair target site.

What is the Zinc-Finger Consortium?

This system was developed by the Zinc-Finger Consortium as an alternative to commercial sources of engineered zinc-finger arrays. A pair of ZFNs, each with three zinc fingers binding to target DNA, are shown introducing a double-strand break, at the FokI domain, depicted in yellow.

What is the function of ZFN enzyme?

Zinc finger nuclease. Zinc-finger nucleases (ZFNs) are artificial restriction enzymes generated by fusing a zinc finger DNA-binding domain to a DNA-cleavage domain.