|Thomson, James - Jim|
Submitted to: Genesis
Publication Type: Peer reviewed journal
Publication Acceptance Date: 8/8/2006
Publication Date: 9/15/2006
Publication URL: www3.interscience.wiley.com/cgi-bin/fulltext/112786688/PDFSTART
Citation: Thomson, J.G., Ow, D.W. 2006. Site-Specific Recombination Systems for the Genetic Manipulation of Eukaryotic Genomes. Genesis 44(10):465-476. Interpretive Summary: This article describes the demonstration of function of seven prokaryotic site-specific recombination systems in the fission yeast. Four of these are dedicated deletion systems, while the remaining three are integration systems that can also perform inversion and deletions. These new recombination systems represent additional genetic tools for the precise manipulation of eukaryotic genomes.
Technical Abstract: Site-specific recombination systems, such as the bacteriophage Cre-lox and yeast FLP-FRT systems, have become valuable tools for the rearrangement of DNA in higher eukaryotes. As a first step to expanding the repertoire of recombination tools, we screened recombination systems derived from the resolvase/invertase family for site-specific recombinase activity in the fission yeast Schizosaccharomyces pombe. Here, we report that seven recombination systems, four from the small serine resolvase subfamily (CinH, ParA, Tn1721 and Tn5053) and three from the large serine resolvase subfamily (Bxb1, TP901-1 and U153) can catalyze site-specific deletion in S. pombe. Those from the large serine resolvase subfamily were also capable of site-specific integration and inversion. In all cases, the recombination events were precise. Functional operation of these recombination systems in the fission yeast holds promise that they may be further developed as recombination tools for the site-specific rearrangement of plant and animal genomes.