ArticleNucleic acids research2010
Directed evolution of recombinase specificity by split gene reassembly.
Article in Nucleic acids research, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
31 citing papers in PubMed.
- Programmable enzymes for targeted gene insertion.Nature reviews. Genetics · 2026Review
- Retargeted serine integrases for one-step, precise integration of large DNA sequences in human cells.Nature biotechnology · 2026Article
- Conformational dynamics promotes disordered regions from function-dispensable to essential in evolved site-specific DNA recombinases.Computational and structural biotechnology journal · 2022Article
- RNA-Guided Recombinase-Cas9 Fusion Targets Genomic DNA Deletion and Integration.The CRISPR journal · 2019Article
- Induction of apoptosis in imatinib sensitive and resistant chronic myeloid leukemia cells by efficient disruption of bcr-abl oncogene with zinc finger nucleases.Journal of experimental & clinical cancer research : CR · 2018Article
- An att site-based recombination reporter system for genome engineering and synthetic DNA assembly.BMC biotechnology · 2017Article
- A programmable Cas9-serine recombinase fusion protein that operates on DNA sequences in mammalian cells.Nucleic acids research · 2016Article
- A systematic survey of the Cys2His2 zinc finger DNA-binding landscape.Nucleic acids research · 2015Article
- Recent advances in developing molecular tools for targeted genome engineering of mammalian cells.BMB reports · 2015Review
- Article
- Synthetic zinc finger proteins: the advent of targeted gene regulation and genome modification technologies.Accounts of chemical research · 2014Article
- Enhancing the specificity of recombinase-mediated genome engineering through dimer interface redesign.Journal of the American Chemical Society · 2014Article
- Expanding the zinc-finger recombinase repertoire: directed evolution and mutational analysis of serine recombinase specificity determinants.Nucleic acids research · 2014Article
- Deep sequencing of large library selections allows computational discovery of diverse sets of zinc fingers that bind common targets.Nucleic acids research · 2014Article
- Expanding the scope of site-specific recombinases for genetic and metabolic engineering.Biotechnology and bioengineering · 2014Review
- ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering.Trends in biotechnology · 2013Review
- A comprehensive approach to zinc-finger recombinase customization enables genomic targeting in human cells.Nucleic acids research · 2013Article
- Mutants of Cre recombinase with improved accuracy.Nature communications · 2013Article
- Chimeric TALE recombinases with programmable DNA sequence specificity.Nucleic acids research · 2012Article
- Chimeric piggyBac transposases for genomic targeting in human cells.Nucleic acids research · 2012Article
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
The engineering of new enzymes that efficiently and specifically modify DNA sequences is necessary for the development of enhanced gene therapies and genetic studies. To address this need, we developed a robust strategy for evolving site-specific recombinases with novel substrate specificities. In this system, recombinase variants are selected for activity on new substrates based on enzyme-mediated reassembly of the gene encoding beta-lactamase that confers ampicillin resistance to Escherichia coli. This stringent evolution method was used to alter the specificities of catalytic domains in the context of a modular zinc finger-recombinase fusion protein. Gene reassembly was detectable over several orders of magnitude, which allowed for tunable selectivity and exceptional sensitivity. Engineered recombinases were evolved to react with sequences from the human genome with only three rounds of selection. Many of the evolved residues, selected from a randomly-mutated library, were conserved among other members of this family of recombinases. This enhanced evolution system will translate recombinase engineering and genome editing into a practical and expedient endeavor for academic, industrial and clinical applications.
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Registered trials
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.