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Harnessing a high cargo-capacity transposon for genetic applications in vertebrates. PLoS Genet 2006 Nov 10;2(11):e169

Date

11/14/2006

Pubmed ID

17096595

Pubmed Central ID

PMC1635535

DOI

10.1371/journal.pgen.0020169

Scopus ID

2-s2.0-33751352744   177 Citations

Abstract

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.

Author List

Balciunas D, Wangensteen KJ, Wilber A, Bell J, Geurts A, Sivasubbu S, Wang X, Hackett PB, Largaespada DA, McIvor RS, Ekker SC

Author

Aron Geurts PhD Associate Professor in the Physiology department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Base Sequence
Cells, Cultured
DNA Transposable Elements
Disease Models, Animal
Exons
Gene Expression
Gene Transfer Techniques
Humans
Mice
Molecular Sequence Data
Tyrosinemias
Zebrafish
jenkins-FCD Prod-398 336d56a365602aa89dcc112f077233607d6a5abc