ArticleJournal of molecular biology2010
Directed evolution of an enhanced and highly efficient FokI cleavage domain for zinc finger nucleases.
Article in Journal of molecular biology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 112 papers.
What it found
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Who cites it
112 citing papers in PubMed.
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- Lipid nanoparticle delivery of TALEN mRNA targeting LPA causes gene disruption and plasma lipoprotein(a) reduction in transgenic mice.Molecular therapy : the journal of the American Society of Gene Therapy · 2025Article
- The generation of novel epialleles in plants: the prospective behind re-shaping the epigenome.Frontiers in plant science · 2025Review
- Genome editing in Sub-Saharan Africa: a game-changing strategy for climate change mitigation and sustainable agriculture.GM crops & food · 2024Review
- Synthetic Gene Circuits for Regulation of Next-Generation Cell-Based Therapeutics.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Review
- A Photoresponsive Homing Endonuclease for Programmed DNA Cleavage.ACS synthetic biology · 2024Article
- Nanoparticle-mediated delivery of non-viral gene editing technology to the brain.Progress in neurobiology · 2024Review
- CRISPR-Cas9 Direct Fusions for Improved Genome Editing via Enhanced Homologous Recombination.International journal of molecular sciences · 2023Review
- Targeted DNA Demethylation: Vectors, Effectors and Perspectives.Biomedicines · 2023Review
- Gene-Modified Stem Cells for Spinal Cord Injury: a Promising Better Alternative Therapy.Stem cell reviews and reports · 2022Review
- Gold Nanoparticle-Mediated Gene Therapy.Cancers · 2022Review
- Cytoplasmic Injection of Zygotes to Genome Edit Naturally Occurring Sequence Variants Into Bovine Embryos.Frontiers in genetics · 2022Article
- Harnessing the power of directed evolution to improve genome editing systems.Current opinion in chemical biology · 2021Review
- Genome Editing Technology and Its Application Potentials in the Industrial Filamentous FungusJournal of fungi (Basel, Switzerland) · 2021Review
- Improved Specificity and Safety of Anti-Hepatitis B Virus TALENs Using Obligate HeterodimericViruses · 2021Article
- Genome engineering and disease modelingWorld journal of stem cells · 2021Review
- CRISPR/Cas systems: opportunities and challenges for crop breeding.Plant cell reports · 2021Review
52 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
Zinc finger nucleases (ZFNs) are powerful tools for gene therapy and genetic engineering. The high specificity and affinity of these chimeric enzymes are based on custom-designed zinc finger proteins (ZFPs). To improve the performance of existing ZFN technology, we developed an in vivo evolution-based approach to improve the efficacy of the FokI cleavage domain (FCD). After multiple rounds of cycling mutagenesis and DNA shuffling, a more efficient nuclease variant (Sharkey) was generated. In vivo analyses indicated that Sharkey is >15-fold more active than wild-type FCD on a diverse panel of cleavage sites. Further, a mammalian cell-based assay showed a three to sixfold improvement in targeted mutagenesis for ZFNs containing derivatives of the Sharkey cleavage domain. We also identified mutations that impart sequence specificity to the FCD that might be utilized in future studies to further refine ZFNs through cooperative specificity. In addition, Sharkey was observed to enhance the cleavage profiles of previously published and newly selected heterodimer ZFN architectures. This enhanced and highly efficient cleavage domain will aid in a variety of ZFN applications in medicine and biology.
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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.