Engineering of a target site-specific recombinase by a combined evolution- and structure-guided approach

Research output: Contribution to journalResearch articleContributedpeer-review



Site-specific recombinases (SSRs) can perform DNA rearrangements, including deletions, inversions and translocations when their naive target sequences are placed strategically into the genome of an organism. Hence, in order to employ SSRs in heterologous hosts, their target sites have to be introduced into the genome of an organism before the enzyme can be practically employed. Engineered SSRs hold great promise for biotechnology and advanced biomedical applications, as they promise to extend the usefulness of SSRs to allow efficient and specific recombination of pre-existing, natural genomic sequences. However, the generation of enzymes with desired properties remains challenging. Here, we use substrate-linked directed evolution in combination with molecular modeling to rationally engineer an efficient and specific recombinase (sTre) that readily and specifically recombines a sequence present in the HIV-1 genome. We elucidate the role of key residues implicated in the molecular recognition mechanism and we present a rationale for sTre's enhanced specificity. Combining evolutionary and rational approaches should help in accelerating the generation of enzymes with desired properties for use in biotechnology and biomedicine.


Original languageEnglish
Pages (from-to)2394-403
Number of pages10
JournalNucleic acids research
Issue number4
Publication statusPublished - 1 Feb 2013

External IDs

PubMedCentral PMC3575804
Scopus 84876397726


Sustainable Development Goals


  • Attachment Sites, Microbiological, DNA/chemistry, Directed Molecular Evolution/methods, Models, Molecular, Protein Binding, Recombinases/chemistry

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