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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...

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Autores principales: 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
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2006
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.
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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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