HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Summary Full Text Full Text (PDF) All Versions of this Article: fj.05-5485fjev1 20/11/1880    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Maragathavally, K. J. Articles by Coates, C. J. Search for Related Content PubMed PubMed Citation Articles by Maragathavally, K. J. Articles by Coates, C. J.

Chimeric Mos1 and piggyBac transposases result in site-directed integration

Department of Entomology, Texas A&M University, College Station, Texas, USA; and
^* Department of Radiology, Medical College of Georgia, Augusta, Georgia, USA

¹Correspondence: Department of Entomology, Texas A&M University, College Station, Texas, USA. E-mail: ccoates{at}tamu.edu

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 NH₂ 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.—Maragathavally, K. J., Kaminski, J. M., Coates, C. J. Chimeric Mos1 and piggyBac transposases result in site-directed integration.

This article has been cited by other articles:

				N. Sharma, B. Moldt, T. Dalsgaard, T. G. Jensen, and J. G. Mikkelsen Regulated gene insertion by steroid-induced {Phi}C31 integrase Nucleic Acids Res., June 1, 2008; 36(11): e67 - e67. [Abstract] [Full Text] [PDF]

				M. E. Morales, V. H. Mann, K. J. Kines, G. N. Gobert, M. J. Fraser Jr, B. H. Kalinna, J. M. Correnti, E. J. Pearce, and P. J. Brindley piggyBac transposon mediated transgenesis of the human blood fluke, Schistosoma mansoni FASEB J, November 1, 2007; 21(13): 3479 - 3489. [Abstract] [Full Text] [PDF]

				S. C.-Y. Wu, Y.-J. J. Meir, C. J. Coates, A. M. Handler, P. Pelczar, S. Moisyadi, and J. M. Kaminski From the Cover: piggyBac is a flexible and highly active transposon as compared to Sleeping Beauty, Tol2, and Mos1 in mammalian cells PNAS, October 10, 2006; 103(41): 15008 - 15013. [Abstract] [Full Text] [PDF]

				C. Feschotte The piggyBac transposon holds promise for human gene therapy PNAS, October 10, 2006; 103(41): 14981 - 14982. [Full Text] [PDF]