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Harnessing a High Cargo-Capacity Transposon for Genetic Applications in Vertebrates
Viruses and transposons are efficient tools for permanently delivering foreign DNA into vertebrate genomes but exhibit diminished activity when cargo exceeds 8 kilobases (kb). This size restriction limits their molecular genetic and biotechnological utility, such as numerous therapeutically relevant...
Autores principales: | , , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Public Library of Science
2006
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1635535/ https://www.ncbi.nlm.nih.gov/pubmed/17096595 http://dx.doi.org/10.1371/journal.pgen.0020169 |
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author | Balciunas, Darius Wangensteen, Kirk J Wilber, Andrew Bell, Jason Geurts, Aron Sivasubbu, Sridhar Wang, Xin Hackett, Perry B Largaespada, David A McIvor, R. Scott Ekker, Stephen C |
author_facet | Balciunas, Darius Wangensteen, Kirk J Wilber, Andrew Bell, Jason Geurts, Aron Sivasubbu, Sridhar Wang, Xin Hackett, Perry B Largaespada, David A McIvor, R. Scott Ekker, Stephen C |
author_sort | Balciunas, Darius |
collection | PubMed |
description | Viruses and transposons are efficient tools for permanently delivering foreign DNA into vertebrate genomes but exhibit diminished activity when cargo exceeds 8 kilobases (kb). This size restriction limits their molecular genetic and biotechnological utility, such as numerous therapeutically relevant genes that exceed 8 kb in size. Furthermore, a greater payload capacity vector would accommodate more sophisticated cis cargo designs to modulate the expression and mutagenic risk of these molecular therapeutics. We show that the Tol2 transposon can efficiently integrate DNA sequences larger than 10 kb into human cells. We characterize minimal sequences necessary for transposition (miniTol2) in vivo in zebrafish and in vitro in human cells. Both the 8.5-kb Tol2 transposon and 5.8-kb miniTol2 engineered elements readily function to revert the deficiency of fumarylacetoacetate hydrolase in an animal model of hereditary tyrosinemia type 1. Together, Tol2 provides a novel nonviral vector for the delivery of large genetic payloads for gene therapy and other transgenic applications. |
format | Text |
id | pubmed-1635535 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2006 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-16355352006-11-29 Harnessing a High Cargo-Capacity Transposon for Genetic Applications in Vertebrates Balciunas, Darius Wangensteen, Kirk J Wilber, Andrew Bell, Jason Geurts, Aron Sivasubbu, Sridhar Wang, Xin Hackett, Perry B Largaespada, David A McIvor, R. Scott Ekker, Stephen C PLoS Genet Research Article Viruses and transposons are efficient tools for permanently delivering foreign DNA into vertebrate genomes but exhibit diminished activity when cargo exceeds 8 kilobases (kb). This size restriction limits their molecular genetic and biotechnological utility, such as numerous therapeutically relevant genes that exceed 8 kb in size. Furthermore, a greater payload capacity vector would accommodate more sophisticated cis cargo designs to modulate the expression and mutagenic risk of these molecular therapeutics. We show that the Tol2 transposon can efficiently integrate DNA sequences larger than 10 kb into human cells. We characterize minimal sequences necessary for transposition (miniTol2) in vivo in zebrafish and in vitro in human cells. Both the 8.5-kb Tol2 transposon and 5.8-kb miniTol2 engineered elements readily function to revert the deficiency of fumarylacetoacetate hydrolase in an animal model of hereditary tyrosinemia type 1. Together, Tol2 provides a novel nonviral vector for the delivery of large genetic payloads for gene therapy and other transgenic applications. Public Library of Science 2006-11 2006-11-10 /pmc/articles/PMC1635535/ /pubmed/17096595 http://dx.doi.org/10.1371/journal.pgen.0020169 Text en © 2006 Balciunas et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Balciunas, Darius Wangensteen, Kirk J Wilber, Andrew Bell, Jason Geurts, Aron Sivasubbu, Sridhar Wang, Xin Hackett, Perry B Largaespada, David A McIvor, R. Scott Ekker, Stephen C Harnessing a High Cargo-Capacity Transposon for Genetic Applications in Vertebrates |
title | Harnessing a High Cargo-Capacity Transposon for Genetic Applications in Vertebrates |
title_full | Harnessing a High Cargo-Capacity Transposon for Genetic Applications in Vertebrates |
title_fullStr | Harnessing a High Cargo-Capacity Transposon for Genetic Applications in Vertebrates |
title_full_unstemmed | Harnessing a High Cargo-Capacity Transposon for Genetic Applications in Vertebrates |
title_short | Harnessing a High Cargo-Capacity Transposon for Genetic Applications in Vertebrates |
title_sort | harnessing a high cargo-capacity transposon for genetic applications in vertebrates |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1635535/ https://www.ncbi.nlm.nih.gov/pubmed/17096595 http://dx.doi.org/10.1371/journal.pgen.0020169 |
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