New Insights into Host Factor Requirements for Prokaryotic %-Recombinase-mediated Reactions in Mammalian Cells

Diaz, Vicente M.; Servert, Pilar; Priéto, Ignacio; González, Manuel A.; Martı́nez-A, Carlos; Alonso, Juan C.; Bernad, António

doi:10.1074/jbc.m011725200

Cited by 21 publications

(2 citation statements)

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“…Thus, unidirectional systems can potentially be more efficient than bi-directional systems for inversion, translocation and integration reactions. The second type of uni-directional recombination system is the excisiononly type in which ParA-MRS and b-six (Diaz et al 2001) are members. These excision-only systems have identical recombination sites, generally known as resolution (res) sites.…”

Section: Introductionmentioning

confidence: 99%

ParA resolvase catalyzes site-specific excision of DNA from the Arabidopsis genome

et al. 2008

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The small serine resolvase ParA from bacterial plasmids RK2 and RP4 catalyzes the recombination of two identical 133 bp recombination sites known as MRS. Previously, we reported that ParA is active in the fission yeast Schizosaccharomyces pombe. In this work, the parA recombinase gene was placed under the control of the Arabidopsis OXS3 promoter and introduced into Arabidopsis lines harboring a chromosomally integrated MRS-flanked target. The ParA recombinase excised the MRS-flanked DNA and the excision event was detected in subsequent generations in the absence of ParA, indicating germinal transmission of the excision event. The precise site-specific deletion by the ParA recombination system in planta demonstrates that the ParA recombinase can be used to remove transgenic DNA, such as selectable markers or other introduced transgenes that are no longer desired in the final product.

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Section: Introductionmentioning

confidence: 99%

ParA resolvase catalyzes site-specific excision of DNA from the Arabidopsis genome

et al. 2008

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“…The small serine sub-family contains β- six (Diaz et al 2001), γδ- res (Schwikardi and Droge 2000), CinH- RS2 (Kholodii 2001; Thomson and Ow 2006) and ParA- MRS (Gerlitz et al 1990; Thomson et al 2009), where β, γδ, CinH and ParA are small serine recombinases, and six, res , RS2 and MRS are the respective DNA recognition sites. While recombination mediated by these small serine recombinases (a.k.a.…”

Section: Recombinase Types and Their Modes Of Actionmentioning

confidence: 99%

Recombinase technology: applications and possibilities

et al. 2010

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The use of recombinases for genomic engineering is no longer a new technology. In fact, this technology has entered its third decade since the initial discovery that recombinases function in heterologous systems (Sauer in Mol Cell Biol 7(6):2087–2096, 1987). The random insertion of a transgene into a plant genome by traditional methods generates unpredictable expression patterns. This feature of transgenesis makes screening for functional lines with predictable expression labor intensive and time consuming. Furthermore, an antibiotic resistance gene is often left in the final product and the potential escape of such resistance markers into the environment and their potential consumption raises consumer concern. The use of site-specific recombination technology in plant genome manipulation has been demonstrated to effectively resolve complex transgene insertions to single copy, remove unwanted DNA, and precisely insert DNA into known genomic target sites. Recombinases have also been demonstrated capable of site-specific recombination within non-nuclear targets, such as the plastid genome of tobacco. Here, we review multiple uses of site-specific recombination and their application toward plant genomic engineering. We also provide alternative strategies for the combined use of multiple site-specific recombinase systems for genome engineering to precisely insert transgenes into a pre-determined locus, and removal of unwanted selectable marker genes.

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Site‐specific recombination systems for the genetic manipulation of eukaryotic genomes

Thomson

2006

Genesis

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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.

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New Insights into Host Factor Requirements for Prokaryotic %-Recombinase-mediated Reactions in Mammalian Cells

Cited by 21 publications

References 50 publications

ParA resolvase catalyzes site-specific excision of DNA from the Arabidopsis genome

ParA resolvase catalyzes site-specific excision of DNA from the Arabidopsis genome

Recombinase technology: applications and possibilities

Site‐specific recombination systems for the genetic manipulation of eukaryotic genomes

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