ArticleNucleic acids research2009
Creation of a type IIS restriction endonuclease with a long recognition sequence.
Article in Nucleic acids research, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 32 citations in OpenAlex.
- An Update on the Application of CRISPR Technology in Clinical Practice.Molecular biotechnology · 2024Review
- Promiscuous DNA cleavage by HpyAII endonuclease is modulated by the HNH catalytic residues.Bioscience reports · 2020Article
- Recent advances in the CRISPR genome editing tool set.Experimental & molecular medicine · 2019Review
- Overview of Post Cohen-Boyer Methods for Single Segment Cloning and for Multisegment DNA Assembly.Current protocols in molecular biology · 2016Article
- Restriction site free cloning (RSFC) plasmid family for seamless, sequence independent cloning in Pichia pastoris.Microbial cell factories · 2015Article
- iBrick: a new standard for iterative assembly of biological parts with homing endonucleases.PloS one · 2014Article
- The MASTER (methylation-assisted tailorable ends rational) ligation method for seamless DNA assembly.Nucleic acids research · 2013Article
- Site- and strand-specific nicking of DNA by fusion proteins derived from MutH and I-SceI or TALE repeats.Nucleic acids research · 2013Article
- Creating highly specific nucleases by fusion of active restriction endonucleases and catalytically inactive homing endonucleases.Nucleic acids research · 2012Article
- Bifunctional TaqII restriction endonuclease: redefining the prototype DNA recognition site and establishing the Fidelity Index for partial cleaving.BMC biochemistry · 2011Article
- Meganucleases and other tools for targeted genome engineering: perspectives and challenges for gene therapy.Current gene therapy · 2011Review
Corrections and comments
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Type IIS restriction endonucleases cleave DNA outside their recognition sequences, and are therefore particularly useful in the assembly of DNA from smaller fragments. A limitation of type IIS restriction endonucleases in assembly of long DNA sequences is the relative abundance of their target sites. To facilitate ligation-based assembly of extremely long pieces of DNA, we have engineered a new type IIS restriction endonuclease that combines the specificity of the homing endonuclease I-SceI with the type IIS cleavage pattern of FokI. We linked a non-cleaving mutant of I-SceI, which conveys to the chimeric enzyme its specificity for an 18-bp DNA sequence, to the catalytic domain of FokI, which cuts DNA at a defined site outside the target site. Whereas previously described chimeric endonucleases do not produce type IIS-like precise DNA overhangs suitable for ligation, our chimeric endonuclease cleaves double-stranded DNA exactly 2 and 6 nt from the target site to generate homogeneous, 5', four-base overhangs, which can be ligated with 90% fidelity. We anticipate that these enzymes will be particularly useful in manipulation of DNA fragments larger than a thousand bases, which are very likely to contain target sites for all natural type IIS restriction endonucleases.
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