The Functionality of Minimal PiggyBac Transposons in Mammalian Cells
Minimal piggyBac vectors are a modified single-plasmid version of the classical piggyBac delivery system that can be used for stable transgene integration. These vectors have a truncated terminal domain in the delivery cassette and thus, integrate significantly less flanking transposon DNA into host...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2016-01-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S216225311730094X |
| _version_ | 1824030992459890688 |
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| author | Boris Troyanovsky Vira Bitko Viktor Pastukh Brian Fouty Victor Solodushko |
| author_facet | Boris Troyanovsky Vira Bitko Viktor Pastukh Brian Fouty Victor Solodushko |
| author_sort | Boris Troyanovsky |
| collection | DOAJ |
| description | Minimal piggyBac vectors are a modified single-plasmid version of the classical piggyBac delivery system that can be used for stable transgene integration. These vectors have a truncated terminal domain in the delivery cassette and thus, integrate significantly less flanking transposon DNA into host cell chromatin than classical piggyBac vectors. Herein, we test various characteristics of this modified transposon. The integration efficiency of minimal piggyBac vectors was inversely related to the size of both the transposon and the entire plasmid, but inserts as large as 15 kb were efficiently integrated. Open and super-coiled vectors demonstrated the same integration efficiency while DNA methylation decreased the integration efficiency and silenced the expression of previously integrated sequences in some cell types. Importantly, the incidence of plasmid backbone integration was not increased above that seen in nontransposon control vectors. In BALB/c mice, we demonstrated prolonged expression of two transgenes (intracellular mCherry and secretable Gaussia luciferase) when delivered by the minimal piggyBac that resulted in a more sustained antibody production against the immunogenic luciferase than when delivered by a transient (nontransposon) vector plasmid. We conclude that minimal piggyBac vectors are an effective alternative to other integrative systems for stable DNA delivery in vitro and in vivo. |
| first_indexed | 2024-12-19T18:48:29Z |
| format | Article |
| id | doaj.art-1454676823654ed1b7ddd57d74c45add |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-19T18:48:29Z |
| publishDate | 2016-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-1454676823654ed1b7ddd57d74c45add2022-12-21T20:09:58ZengElsevierMolecular Therapy: Nucleic Acids2162-25312016-01-015C10.1038/mtna.2016.76The Functionality of Minimal PiggyBac Transposons in Mammalian CellsBoris Troyanovsky0Vira Bitko1Viktor Pastukh2Brian Fouty3Victor Solodushko4Department of Pharmacology, University of South Alabama School of Medicine, Mobile, Alabama, USANanoBio Corporation, Ann Arbor, Michigan, USADepartment of Pharmacology, University of South Alabama School of Medicine, Mobile, Alabama, USADepartment of Pharmacology, University of South Alabama School of Medicine, Mobile, Alabama, USADepartment of Pharmacology, University of South Alabama School of Medicine, Mobile, Alabama, USAMinimal piggyBac vectors are a modified single-plasmid version of the classical piggyBac delivery system that can be used for stable transgene integration. These vectors have a truncated terminal domain in the delivery cassette and thus, integrate significantly less flanking transposon DNA into host cell chromatin than classical piggyBac vectors. Herein, we test various characteristics of this modified transposon. The integration efficiency of minimal piggyBac vectors was inversely related to the size of both the transposon and the entire plasmid, but inserts as large as 15 kb were efficiently integrated. Open and super-coiled vectors demonstrated the same integration efficiency while DNA methylation decreased the integration efficiency and silenced the expression of previously integrated sequences in some cell types. Importantly, the incidence of plasmid backbone integration was not increased above that seen in nontransposon control vectors. In BALB/c mice, we demonstrated prolonged expression of two transgenes (intracellular mCherry and secretable Gaussia luciferase) when delivered by the minimal piggyBac that resulted in a more sustained antibody production against the immunogenic luciferase than when delivered by a transient (nontransposon) vector plasmid. We conclude that minimal piggyBac vectors are an effective alternative to other integrative systems for stable DNA delivery in vitro and in vivo.http://www.sciencedirect.com/science/article/pii/S216225311730094Xminimal piggyBac vectorsstable gene deliverytransposon |
| spellingShingle | Boris Troyanovsky Vira Bitko Viktor Pastukh Brian Fouty Victor Solodushko The Functionality of Minimal PiggyBac Transposons in Mammalian Cells Molecular Therapy: Nucleic Acids minimal piggyBac vectors stable gene delivery transposon |
| title | The Functionality of Minimal PiggyBac Transposons in Mammalian Cells |
| title_full | The Functionality of Minimal PiggyBac Transposons in Mammalian Cells |
| title_fullStr | The Functionality of Minimal PiggyBac Transposons in Mammalian Cells |
| title_full_unstemmed | The Functionality of Minimal PiggyBac Transposons in Mammalian Cells |
| title_short | The Functionality of Minimal PiggyBac Transposons in Mammalian Cells |
| title_sort | functionality of minimal piggybac transposons in mammalian cells |
| topic | minimal piggyBac vectors stable gene delivery transposon |
| url | http://www.sciencedirect.com/science/article/pii/S216225311730094X |
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