Site-specific recombinase, R, encoded by yeast plasmid pSR1

Araki, Hideki; Nakanishi, Noriyuki; Evans, Barbara R.; Matsuzaki, Hiroaki; Jayaram, Makkuni; Oshima, Yasuji

doi:10.1016/0022-2836(92)91023-i

Cited by 42 publications

(29 citation statements)

References 34 publications

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“…The weaker properties of FLP, as well as the merits in finding a third efficient recombinase for genome engineering, prompted searches for new tyrosine SSR tools (Araki et al, 1992;Ringrose et al, 1997;Christ et al, 2002 …”

Section: Introductionmentioning

confidence: 99%

Dre recombinase, like Cre, is a highly efficient site-specific recombinase in E. coli, mammalian cells and mice

Anastassiadis

Patsch

et al. 2009

Disease Models &Amp; Mechanisms

269

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SUMMARY Tyrosine site-specific recombinases (SSRs) including Cre and FLP are essential tools for DNA and genome engineering. Cre has long been recognized as the best SSR for genome engineering, particularly in mice. Obtaining another SSR that is as good as Cre will be a valuable addition to the genomic toolbox. To this end, we have developed and validated reagents for the Dre-rox system. These include an Escherichia coli-inducible expression vector based on the temperature-sensitive pSC101 plasmid, a mammalian expression vector based on the CAGGs promoter, a rox-lacZ reporter embryonic stem (ES) cell line based on targeting at the Rosa26 locus, the accompanying Rosa26-rox reporter mouse line, and a CAGGs-Dre deleter mouse line. We also show that a Dre-progesterone receptor shows good ligand-responsive induction properties. Furthermore, we show that there is no crossover recombination between Cre-rox or Dre-loxP. Hence, we add another set of efficient tools to the genomic toolbox, which will enable the development of more sophisticated mouse models for the analysis of gene function and disease.

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Section: Introductionmentioning

confidence: 99%

Dre recombinase, like Cre, is a highly efficient site-specific recombinase in E. coli, mammalian cells and mice

Anastassiadis

Patsch

et al. 2009

Disease Models &Amp; Mechanisms

269

209

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“…34). In particular, they appear to encode site-specific recombinases related to FLP and undergo high-frequency intramolecular recombination between inverted repeat domains both in their native species and in S. cerevisiae (32,(34)(35)(36)(37)(38)(39). However, only the R recombinase has previously been shown to work in a multicellular organism (33), and none has been used in an animal genome.…”

mentioning

confidence: 99%

Multiple new site-specific recombinases for use in manipulating animal genomes

Nern

Pfeiffer

Svoboda

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

160

171

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Site-specific recombinases have been used for two decades to manipulate the structure of animal genomes in highly predictable ways and have become major research tools. However, the small number of recombinases demonstrated to have distinct specificities, low toxicity, and sufficient activity to drive reactions to completion in animals has been a limitation. In this report we show that four recombinases derived from yeast-KD, B2, B3, and R-are highly active and nontoxic in Drosophila and that KD, B2, B3, and the widely used FLP recombinase have distinct target specificities. We also show that the KD and B3 recombinases are active in mice.gene expression | genetic engineering S ite-specific DNA recombinases are widely used in multicellular organisms to manipulate the structure of genomes and, in turn, to control gene expression (for reviews see refs. 1-4). These enzymes, derived from bacteria and fungi, catalyze directionally sensitive DNA exchange reactions between short (30-40 nucleotides) target site sequences that are specific to each recombinase (5). These reactions enable four basic functional modules-excision/insertion, inversion, translocation and cassette exchange-that have been used individually or combined in a wide range of configurations to control gene expression (Fig. 1A).The use of site-specific recombination to manipulate genomes has been limited by the availability of recombinases with high activity, distinct site specificity, and low toxicity. In Drosophila, the most widely used recombinase is FLP, encoded by the Saccharomyces cerevisiae 2-μm plasmid (6). FLP was first shown to work in a heterologous, multicellular organism by Golic and Lindquist in 1989 (7) who demonstrated the excision reaction on chromosomally inserted target sites (FRTs). Since that time FLP/FRT recombination has been widely used in Drosophila in applications based on excision (8) and translocation (9-11).Complex manipulations of genome structure can require the use of more than one of the modules diagrammed in Fig. 1A, or parallel independent implementations of the same module, in a single individual. To accomplish such manipulations, the modules must be implemented with different recombinases that do not recognize each other's target sites. Similarly, a number of powerful methods have been developed for using the excision and inversion reactions to control expression of a transgene specifically in cells where two independent gene expression patterns overlap (2,3,12,13). Such intersectional methods rely on pairs of orthogonal recombinases; see, for example, Fig. 1B. For these reasons, we sought to discover additional recombinases with distinct site-specificity.FLP recombinase has been mutated to recognize altered FRT sites, but some cross-reaction still remains (14, 15). Cre, encoded by the bacteriophage P1, is the most widely used recombinase in mammalian cells (16)(17)(18). Cre functions in Drosophila (19), but exhibits obvious toxicity (20), a problem also observed in mammalian cells (21, 22; reviewed in ref. 23) an...

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“…Using two DNA fragments as probes (probes 1 and 2) for Southern hybridization, we were able to distinguish the relative direction of each transposition. We also introduced RS sequences, namely, recognition sites for recombinase (R protein) from Zygosaccaromyces rouxii (Araki et al,1985, Matsuzaki et al, 1988, Araki et al, 1992 into the T-DNA construct: two RSs in inverted orientation were placed inside and outside dAc-I-RS, respectively, as described previously (Machida et al, 2000). In addition to such specific DNA sequences, we inserted the gene for hygromycin phosphotransferase (HPT) under the control of a 35S promoter into the dAc-I-RS element for selection of transgenic cell lines and detection of dAc-I-RS insertion (Fig.…”

Section: Resultsmentioning

confidence: 99%

The transposition pattern of the Ac element in tobacco cultured cells.

et al. 2001

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We investigated physical distances and directions of transposition of the maize transposable element Ac in tobacco cultured cells. We introduced a T-DNA construct that carried a non-autonomous derivative of Ac (designated dAc-I-RS) that included sites for cleavage by restriction endonuclease MluI. Another cleavage site was also introduced into the T-DNA region outside of the dAc-I-RS transposable element. The tobacco cultured cell line BY-2 was transformed with the T-DNA and several transformed lines that had a single copy of the T-DNA at a different chromosomal location were isolated. These lines were co-cultured with Agrobacterium tumefaciens cells that carried a cDNA for the Ac transposase gene under the control of various promoters. Sublines of cultured cells in which dAc-I-RS had been transposed, were isolated. The genomic DNAs of these sublines were isolated and digested with MluI. Sizes of DNA segments generated by digestion were determined by pulse-field gel electrophoresis. Our results showed that 20 to 70% of transposition events had occurred within several hundreds kilo-base pairs (kb) on the same chromosome. These results demonstrate that the Ac-Ds element preferentially transposed to regions near the original site in a tobacco chromosome. In addition, the present results are an example of asymmetric transposition as demonstrated by the distance of transposition on the chromosome.

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Site-specific recombinase, R, encoded by yeast plasmid pSR1

Cited by 42 publications

References 34 publications

Dre recombinase, like Cre, is a highly efficient site-specific recombinase in E. coli, mammalian cells and mice

Dre recombinase, like Cre, is a highly efficient site-specific recombinase in E. coli, mammalian cells and mice

Multiple new site-specific recombinases for use in manipulating animal genomes

The transposition pattern of the Ac element in tobacco cultured cells.

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