ArticleNucleic acids research2009
Rational engineering of type II restriction endonuclease DNA binding and cleavage specificity.
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 24 papers.
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Who cites it
24 citing papers in PubMed, 49 citations in OpenAlex.
- Recent Advancements for Enhanced Biocatalyst and Biotransformation.Molecular biotechnology · 2026Review
- Decoding and reengineering the promoter specificity of T7-like RNA polymerases based on phage genome sequences.Nucleic acids research · 2025Article
- Structure and rational engineering of the PglX methyltransferase and specificity factor for BREX phage defence.Nature communications · 2024Article
- Article
- Expression and Purification ofFrontiers in microbiology · 2022Article
- Coordination of phage genome degradation versus host genome protection by a bifunctional restriction-modification enzyme visualized by CryoEM.Structure (London, England : 1993) · 2021Article
- Engineering altered protein-DNA recognition specificity.Nucleic acids research · 2018Review
- The third restriction-modification system from Thermus aquaticus YT-1: solving the riddle of two TaqII specificities.Nucleic acids research · 2017Article
- DNA target recognition domains in the Type I restriction and modification systems of Staphylococcus aureus.Nucleic acids research · 2017Article
- Structural insights into DNA sequence recognition by Type ISP restriction-modification enzymes.Nucleic acids research · 2016Article
- Structure of Type IIL Restriction-Modification Enzyme MmeI in Complex with DNA Has Implications for Engineering New Specificities.PLoS biology · 2016Article
- Generation of artificial sequence-specific nucleasesChemical science · 2016Article
- Article
- Type II restriction endonucleases--a historical perspective and more.Nucleic acids research · 2014Review
- Methylome diversification through changes in DNA methyltransferase sequence specificity.PLoS genetics · 2014Article
- The complex methylome of the human gastric pathogen Helicobacter pylori.Nucleic acids research · 2014Article
- Characterization of cleavage intermediate and star sites of RM.Tth111II.Scientific reports · 2014Article
- Type I restriction enzymes and their relatives.Nucleic acids research · 2014Review
- Impact of target site distribution for Type I restriction enzymes on the evolution of methicillin-resistant Staphylococcus aureus (MRSA) populations.Nucleic acids research · 2013Article
- The methylomes of six bacteria.Nucleic acids research · 2012Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The type II restriction endonucleases are indispensible tools for molecular biology. Although enzymes recognizing nearly 300 unique sequences are known, the ability to engineer enzymes to recognize any sequence of choice would be valuable. However, previous attempts to engineer new recognition specificity have met limited success. Here we report the rational engineering of multiple new type II specificities. We recently identified a family of MmeI-like type II endonucleases that have highly similar protein sequences but different recognition specificity. We identified the amino-acid positions within these enzymes that determine position specific DNA base recognition at three positions within their recognition sequences through correlations between their aligned amino-acid residues and aligned recognition sequences. We then altered the amino acids at the identified positions to those correlated with recognition of a desired new base to create enzymes that recognize and cut at predictable new DNA sequences. The enzymes so altered have similar levels of endonuclease activity compared to the wild-type enzymes. Using simple and predictable mutagenesis in this family it is now possible to create hundreds of unique new type II restriction endonuclease specificities. The findings suggest a simple mechanism for the evolution of new DNA specificity in Nature.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.