2012
DOI: 10.1021/bc200480m
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Catalytic Activity Control of Restriction Endonuclease—Triplex Forming Oligonucleotide Conjugates

Abstract: Targeting of individual genes in complex genomes requires endonucleases of extremely high specificity. To direct cleavage at the unique site(s) in the genome, both naturally occurring and artificial enzymes have been developed. These include homing endonucleases, zinc-finger nucleases, transcription activator-like effector nucleases, and restriction or chemical nucleases coupled to a triple-helix forming oligonucleotide (TFO). The desired cleavage has been demonstrated both in vivo and in vitro for several mod… Show more

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Cited by 8 publications
(9 citation statements)
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“…In most HEs the DNA-binding domains are not distinct from the cleavage domains this makes engineering these enzymes more difficult but HEs may have greater selectivity (i.e., specificity) for the intended target site compared with the modular ZFNs (Zeevi et al 2012) or the recently developed transcription activator-like effector nucleases (TALENs) ( Fig. 3d; Schleifman et al 2008;Vainstein et al 2011;Silanskas et al 2012).…”
Section: Future Prospects and Alternative Genomeediting Toolsmentioning
confidence: 99%
See 1 more Smart Citation
“…In most HEs the DNA-binding domains are not distinct from the cleavage domains this makes engineering these enzymes more difficult but HEs may have greater selectivity (i.e., specificity) for the intended target site compared with the modular ZFNs (Zeevi et al 2012) or the recently developed transcription activator-like effector nucleases (TALENs) ( Fig. 3d; Schleifman et al 2008;Vainstein et al 2011;Silanskas et al 2012).…”
Section: Future Prospects and Alternative Genomeediting Toolsmentioning
confidence: 99%
“…Another potential alternative to HEs are the triplex-forming oligonucleotide (TFO) nucleas•es, here a type II RE domain is coupled to a TFO, the latter can be engineered as needed and the oligonucleotide provide a sequence-specific DNA ligand, instead of a protein-based binding domain as seen in HEs, ZFNs, or TALENs ( Fig. 3e; Eisenschmidt et al 2005;Schleifman et al 2008;Silanskas et al 2012). In the original prototype for a TFO nuclease, Eisenschmidt et al (2005) fused a single-chain variant of the PvuII RE (scPvuII) to a TFO to target a specific sequence in vitro.…”
Section: Future Prospects and Alternative Genomeediting Toolsmentioning
confidence: 99%
“…To regulate ZFN expression on transcriptional level, Silanskas et al (2012) fused a ubiquitin moiety or FKBP12 domain to the N-terminus of ZFN respectively. Their strategies involved creating ZFNs with shortened half-lives and then regulating protein levels with small molecules.…”
Section: Discussionmentioning
confidence: 99%
“…This residue works as an enzyme inactivator, meaning that as long as the residue is bound to the enzyme, the enzyme exhibits no or low activity . Once the residue is cleaved upon stimulation, typically irradiation, the enzyme activity is restored . The mechanism of uncaging an enzyme is schematically shown in Figure .…”
Section: Methods For the Regulation Of Enzyme Activitymentioning
confidence: 99%