piggyBac is a flexible and highly active transposon as compared to Sleeping Beauty, Tol2, and Mos1 in mammalian cells
Sareina Chiung Yuan Wu,Yaa-Jyuhn James Meir,Craig J. Coates,Craig J. Coates,Alfred M. Handler,Pawel Pelczar,Stefan Moisyadi,Stefan Moisyadi,Joseph M. Kaminski +8 more
TLDR
The high transposition activity of piggyBac and the flexibility for molecular modification of its transposase suggest the possibility of using it routinely for mammalian transgenesis.Abstract:
A nonviral vector for highly efficient site-specific integration would be desirable for many applications in transgenesis, including gene therapy. In this study we directly compared the genomic integration efficiencies of piggyBac, hyperactive Sleeping Beauty (SB11), Tol2, and Mos1 in four mammalian cell lines. piggyBac demonstrated significantly higher transposition activity in all cell lines whereas Mos1 had no activity. Furthermore, piggyBac transposase coupled to the GAL4 DNA-binding domain retains transposition activity whereas similarly manipulated gene products of Tol2 and SB11 were inactive. The high transposition activity of piggyBac and the flexibility for molecular modification of its transposase suggest the possibility of using it routinely for mammalian transgenesis.read more
Citations
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Non-viral vectors for gene-based therapy
Hao Yin,Rosemary Lynn Kanasty,Ahmed A. Eltoukhy,Arturo J. Vegas,J. Robert Dorkin,Daniel G. Anderson +5 more
TL;DR: The biological barriers to gene delivery in vivo are introduced and recent advances in material sciences, nanotechnology and nucleic acid chemistry that have yielded promising non-viral delivery systems are discussed, some of which are currently undergoing testing in clinical trials.
Journal ArticleDOI
piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells
Knut Woltjen,Iacovos P. Michael,Iacovos P. Michael,Paria Mohseni,Paria Mohseni,Ridham Desai,Ridham Desai,Maria Mileikovsky,Riikka H. Hämäläinen,Rebecca Cowling,Wei Wang,Pentao Liu,Marina Gertsenstein,Keisuke Kaji,Hoon Ki Sung,Andras Nagy,Andras Nagy +16 more
TL;DR: It is shown that the individual PB insertions can be removed from established iPS cell lines, providing an invaluable tool for discovery, and the traceless removal of reprogramming factors joined with viral 2A sequences delivered by a single transposon from murine iPS lines is demonstrated.
Journal ArticleDOI
A hyperactive piggyBac transposase for mammalian applications.
TL;DR: A unique hyperactive piggyBac transposase is generated with 17-fold and ninefold increases in excision and integration, respectively, and its applicability is shown by demonstrating an increased efficiency of generation of transgene-free mouse induced pluripotent stem cells.
Journal ArticleDOI
Tol2: a versatile gene transfer vector in vertebrates
TL;DR: This review describes and discusses how the Tol2 transposon is being applied to transgenic studies in these vertebrates, and possible future applications.
Journal ArticleDOI
Mammalian cell protein expression for biopharmaceutical production
TL;DR: This review focused on the latest advancements in the field, especially in active areas such as expression systems, glycosylation impact factors, and transient gene expression, which plays more important role in biopharmaceutical early development stages.
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Journal ArticleDOI
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TL;DR: The results of an international collaboration to produce a high-quality draft sequence of the mouse genome are reported and an initial comparative analysis of the Mouse and human genomes is presented, describing some of the insights that can be gleaned from the two sequences.
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TL;DR: Sleeping Beauty is an active DNA-transposon system from vertebrates for genetic transformation and insertional mutagenesis, and it mediates precise cut-and-paste transposition in fish as well as in mouse and human cells.
Journal ArticleDOI
Efficient Transposition of the piggyBac (PB) Transposon in Mammalian Cells and Mice
TL;DR: It is shown that piggyBac elements carrying multiple genes can efficiently transpose in human and mouse cell lines and also in mice, providing a first and critical step toward a highly efficient transposon system for a variety of genetic manipulations in mice and other vertebrates.
Journal ArticleDOI
Mammalian mutagenesis using a highly mobile somatic Sleeping Beauty transposon system
TL;DR: Sleeping Beauty transposition can easily be controlled to mutagenize any target tissue and can therefore, in principle, be used to induce many of the cancers affecting humans, including those for which little is known about the aetiology.