ArticleNucleic acids research2005
Developing a programmed restriction endonuclease for highly specific DNA cleavage.
Article in Nucleic acids research, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed.
- Developing a universal multi-epitope protein vaccine candidate for enhanced borna virus pandemic preparedness.Frontiers in immunology · 2024Article
- The past and presence of gene targeting: from chemicals and DNA via proteins to RNA.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2018Review
- From Bioengineering to CRISPR/Cas9 - A Personal Retrospective of 20 Years of Research in Programmable Genome Targeting.Frontiers in genetics · 2018Article
- TALE-PvuII fusion proteins--novel tools for gene targeting.PloS one · 2013Article
- Chromosomal context and epigenetic mechanisms control the efficacy of genome editing by rare-cutting designer endonucleases.Nucleic acids research · 2012Article
- A novel zinc-finger nuclease platform with a sequence-specific cleavage module.Nucleic acids research · 2012Article
- Creating highly specific nucleases by fusion of active restriction endonucleases and catalytically inactive homing endonucleases.Nucleic acids research · 2012Article
- Review
- Meganucleases and other tools for targeted genome engineering: perspectives and challenges for gene therapy.Current gene therapy · 2011Review
- Targeted DNA methylation by a DNA methyltransferase coupled to a triple helix forming oligonucleotide to down-regulate the epithelial cell adhesion molecule.Bioconjugate chemistry · 2010Article
- Molecular scissors under light control.Proceedings of the National Academy of Sciences of the United States of America · 2010Article
- Controlling the enzymatic activity of a restriction enzyme by light.Proceedings of the National Academy of Sciences of the United States of America · 2010Article
- Efficient targeting of a SCID gene by an engineered single-chain homing endonuclease.Nucleic acids research · 2009Article
- Targeted generation of DNA strand breaks using pyrene-conjugated triplex-forming oligonucleotides.Biochemistry · 2008Article
- Expression and purification of BmrI restriction endonuclease and its N-terminal cleavage domain variants.Protein expression and purification · 2008Article
- Purification, crystallization, X-ray diffraction analysis and phasing of an engineered single-chain PvuII restriction endonuclease.Acta crystallographica. Section F, Structural biology and crystallization communications · 2007Article
- Catalytic domain of restriction endonuclease BmrI as a cleavage module for engineering endonucleases with novel substrate specificities.Nucleic acids research · 2007Article
- Chemical modification of Ce(IV)/EDTA-based artificial restriction DNA cutter for versatile manipulation of double-stranded DNA.Nucleic acids research · 2007Article
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Specific cleavage of large DNA molecules at few sites, necessary for the analysis of genomic DNA or for targeting individual genes in complex genomes, requires endonucleases of extremely high specificity. Restriction endonucleases (REase) that recognize DNA sequences of 4-8 bp are not sufficiently specific for this purpose. In principle, the specificity of REases can be extended by fusion to sequence recognition modules, e.g. specific DNA-binding domains or triple-helix forming oligonucleotides (TFO). We have chosen to extend the specificity of REases using TFOs, given the combinatorial flexibility this fusion offers in addressing a short, yet precisely recognized restriction site next to a defined triple-helix forming site (TFS). We demonstrate here that the single chain variant of PvuII (scPvuII) covalently coupled via the bifunctional cross-linker N-(gamma-maleimidobutryloxy) succinimide ester to a TFO (5'-NH2-[CH2](6 or 12)-MPMPMPMPMPPPPPPT-3', with M being 5-methyl-2'-deoxycytidine and P being 5-[1-propynyl]-2'-deoxyuridine), cleaves DNA specifically at the recognition site of PvuII (CAGCTG) if located in a distance of approximately one helical turn to a TFS (underlined) complementary to the TFO ('addressed' site: 5'-TTTTTTTCTCTCTCTCN(approximately 10)CAGCTG-3'), leaving 'unaddressed' PvuII sites intact. The preference for cleavage of an 'addressed' compared to an 'unaddressed' site is >1000-fold, if the cleavage reaction is initiated by addition of Mg2+ ions after preincubation of scPvuII-TFO and substrate in the absence of Mg2+ ions to allow triple-helix formation before DNA cleavage. Single base pair substitutions in the TFS prevent addressed DNA cleavage by scPvuII-TFO.
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