Chimeric Mos1 and piggyBac transposases result in site-directed integration.

K. J. Maragathavally, J. M. Kaminski, C. J. Coates

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Genetic transformation systems based on Mos1 and piggyBac transposable elements are used to achieve stable chromosomal integration. However, integration sites are randomly distributed in the genome and transgene expression can be influenced by position effects. We developed a novel technology that utilizes chimeric transposases to direct integration into specific sites on a target DNA molecule. The Gal4 DNA binding domain was fused to the NH(2) terminus of the Mos1 and piggyBac transposases and a target plasmid was created that contained upstream activating sequences (UAS), to which the Gal4 DBD binds with high affinity. The transpositional activity of the Gal4-Mos1 transposase was 12.7-fold higher compared to controls where the Gal4-UAS interaction was absent and 96% of the recovered transposition products were identical, with integration occurring at the same TA site. In a parallel experiment, a Gal4-piggyBac transposase resulted in an 11.6-fold increase in transpositional activity compared to controls, with 67% of the integrations occurring at a single TTAA site. This technology has the potential to minimize nonspecific integration events that may result in insertional mutagenesis and reduced fitness. Site-directed integration will be advantageous to the manipulation of genomes, study of gene function, and for the development of gene therapy techniques.

Original languageEnglish (US)
Pages (from-to)1880-1882
Number of pages3
JournalThe FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Volume20
Issue number11
DOIs
StatePublished - Sep 1 2006

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Transposases
Genome
Technology
Genetic Transformation
DNA Transposable Elements
Insertional Mutagenesis
DNA
Transgenes
Genetic Therapy
Plasmids
Genes
Gene therapy
Mutagenesis
mariner transposases
Molecules

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Chimeric Mos1 and piggyBac transposases result in site-directed integration. / Maragathavally, K. J.; Kaminski, J. M.; Coates, C. J.

In: The FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Vol. 20, No. 11, 01.09.2006, p. 1880-1882.

Research output: Contribution to journalArticle

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