ArticleProceedings of the National Academy of Sciences of the United States of America2013
piggyBac transposase tools for genome engineering.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 149 papers.
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Who cites it
149 citing papers in PubMed.
- Mining and engineering of activeSynthetic and systems biotechnology · 2027Article
- Progresses of transposon research inSynthetic and systems biotechnology · 2026Review
- Discovery and protein language model-guided design of hyperactive transposases.Nature biotechnology · 2026Article
- Transposase-Assisted Donor Tethering Boosts Large-Fragment HDR in Plants.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- A closed-loop cell therapy engineered to autonomously secrete activin A inhibitor protects from fibrodysplasia ossificans progressiva.iScience · 2026Article
- Structure-based recasting of a mammalian DNA transpososome as an obligate heterodimer.bioRxiv : the preprint server for biology · 2026Article
- Generation of transgenic pigs with targeted insertion of a wildtype copy of human PKD2 gene.Molecular biology reports · 2026Article
- Review
- Tunable gene control via RNA splicing with a clinically approved small molecule.Nature communications · 2026Article
- Improved vector toolkit for genome writing in mammalian cells.bioRxiv : the preprint server for biology · 2026Article
- A closed-loop cell therapy engineered to autonomously secrete Activin A inhibitor protects from fibrodysplasia ossificans progressiva.bioRxiv : the preprint server for biology · 2026Article
- Specific pathogen free ten gene-edited donor pigs for xenotransplantation.Protein & cell · 2025Article
- Gene-sized DNA insertion at genomic safe harbors in human cells using a site-directed transposase.Nucleic acids research · 2025Article
- MultiSite Assembly of Gateway Induced Clones (MAGIC): a flexible cloning toolbox for use in vertebrate model systems.Development (Cambridge, England) · 2025Article
- Article
- Optimizing Lentiviral Vector Production: Insights Into PiggyBac Transposase and Concatemeric Array Strategies.Biotechnology journal · 2025Article
- SunTag-PE: a modular prime editing system enables versatile and efficient genome editing.Communications biology · 2025Article
- Dynamic co-evolution of transposable elements and the piRNA pathway in African cichlid fishes.Genome biology · 2025Article
- biGMamAct: efficient CRISPR/Cas9-mediated docking of large functional DNA cargoes at theSynthetic biology (Oxford, England) · 2025Article
- CAR-NK cells: harnessing the power of natural killers for advanced cancer therapy.Frontiers in immunology · 2025Review
89 more citing papers are in PubMed but not listed here.
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Authors and funding
13 authors.
Funding
Abstract
The transposon piggyBac is being used increasingly for genetic studies. Here, we describe modified versions of piggyBac transposase that have potentially wide-ranging applications, such as reversible transgenesis and modified targeting of insertions. piggyBac is distinguished by its ability to excise precisely, restoring the donor site to its pretransposon state. This characteristic makes piggyBac useful for reversible transgenesis, a potentially valuable feature when generating induced pluripotent stem cells without permanent alterations to genomic sequence. To avoid further genome modification following piggyBac excision by reintegration, we generated an excision competent/integration defective (Exc(+)Int(-)) transposase. Our findings also suggest the position of a target DNA-transposase interaction. Another goal of genome engineering is to develop reagents that can guide transgenes to preferred genomic regions. Others have shown that piggyBac transposase can be active when fused to a heterologous DNA-binding domain. An Exc(+)Int(-) transposase, the intrinsic targeting of which is defective, might also be a useful intermediate in generating a transposase whose integration activity could be rescued and redirected by fusion to a site-specific DNA-binding domain. We show that fusion to two designed zinc finger proteins rescued the Int(-) phenotype. Successful guided transgene integration into genomic DNA would have broad applications to gene therapy and molecular genetics. Thus, an Exc(+)Int(-) transposase is a potentially useful reagent for genome engineering and provides insight into the mechanism of transposase-target DNA interaction.
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