Harnessing a high cargo-capacity transposon for genetic applications in vertebrates. PLoS Genet 2006 Nov 10;2(11):e169
Date
11/14/2006Pubmed ID
17096595Pubmed Central ID
PMC1635535DOI
10.1371/journal.pgen.0020169Scopus ID
2-s2.0-33751352744 (requires institutional sign-in at Scopus site) 261 CitationsAbstract
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 SCAuthor
Aron Geurts PhD Professor in the Physiology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsBase Sequence
Cells, Cultured
DNA Transposable Elements
Disease Models, Animal
Exons
Gene Expression
Gene Transfer Techniques
Humans
Mice
Molecular Sequence Data
Tyrosinemias
Zebrafish
