Submitted to: Trends in Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/11/2004
Publication Date: 12/1/2004
Citation: Srivastava, V., Ow, D.W. 2004. Marker-free site-specific gene integration in plants. Trends in Biotechnology, Trends in Biotechnology, 22(12):627-629. Interpretive Summary: For nearly 15 years, the use of site-specific recombination systems in plants has focused on the deletion or integration of DNA. Each of these applications offers practical solutions to the two problems in biotechnology: the presence of the unneeded DNA in transgene locus and variation of the locus structure between independent transgenic lines. Given that each of these separate applications is becoming more practical for commercial use, it is time to consider combining them into an integrated technology. Here, we describe a strategy for a 'combined step' method that makes use of two site-specific recombination systems, one for integrating the DNA and a second for removing sequences that are not needed after DNA transfer. This strategy is based on published data, and exemplifies the use of Cre-lox, FLP-FRT, or R-RS systems.
Technical Abstract: This article describes how one site-specific recombination system may be used to direct the integration of DNA precisely into a defined chromosome location of the plant genome, while a second site-specific recombination system may be used to remove the unneeded marker gene after successful site-specific integration.