ArticlePlant physiology2007
Stable recombinase-mediated cassette exchange in Arabidopsis using Agrobacterium tumefaciens.
Article in Plant physiology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Development of an efficient genome editing system using recombinase-mediated cassette exchange inSynthetic and systems biotechnology · 2026Article
- Sequence modification on demand: search and replace tools for precise gene editing in plants.Transgenic research · 2021Review
- Targeted DNA insertion in plants.Proceedings of the National Academy of Sciences of the United States of America · 2021Review
- CRAGE enables rapid activation of biosynthetic gene clusters in undomesticated bacteria.Nature microbiology · 2019Article
- High efficiency Agrobacterium-mediated site-specific gene integration in maize utilizing the FLP-FRT recombination system.Plant biotechnology journal · 2019Article
- Construction of Marker-Free Genetically Modified Maize Using a Heat-Inducible Auto-Excision Vector.Genes · 2019Article
- Progress of targeted genome modification approaches in higher plants.Plant cell reports · 2016Review
- Targeted gene exchange in plant cells mediated by a zinc finger nuclease double cut.Plant biotechnology journal · 2016Article
- Gene stacking by recombinases.Plant biotechnology journal · 2016Review
- Recombinase-Mediated Cassette Exchange (RMCE)-in Reporter Cell Lines as an Alternative to the Flp-in System.PloS one · 2016Article
- Gene stacking in plant cell using recombinases for gene integration and nucleases for marker gene deletion.BMC biotechnology · 2015Article
- Cas9-Guide RNA Directed Genome Editing in Soybean.Plant physiology · 2015Article
- Engineering of plant chromosomes.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2015Review
- Site-specific cassette exchange systems in the Aedes aegypti mosquito and the Plutella xylostella moth.PloS one · 2015Article
- Minichromosomes and artificial chromosomes in Arabidopsis.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2014Review
- Engineering complex biological systems in bacteria through recombinase-assisted genome engineering.Nature protocols · 2014Article
- Improved FLP recombinase, FLPe, efficiently removes marker gene from transgene locus developed by Cre-lox mediated site-specific gene integration in rice.Molecular biotechnology · 2011Article
- Recombinase technology: applications and possibilities.Plant cell reports · 2011Review
- Stacking multiple transgenes at a selected genomic site via repeated recombinase-mediated DNA cassette exchanges.Plant physiology · 2010Article
- Site-specific integration of transgenes in soybean via recombinase-mediated DNA cassette exchange.Plant physiology · 2009Article
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Site-specific integration is an attractive method for the improvement of current transformation technologies aimed at the production of stable transgenic plants. Here, we present a Cre-based targeting strategy in Arabidopsis (Arabidopsis thaliana) using recombinase-mediated cassette exchange (RMCE) of transferred DNA (T-DNA) delivered by Agrobacterium tumefaciens. The rationale for effective RMCE is the precise exchange of a genomic and a replacement cassette both flanked by two heterospecific lox sites that are incompatible with each other to prevent unwanted cassette deletion. We designed a strategy in which the coding region of a loxP/lox5171-flanked bialaphos resistance (bar) gene is exchanged for a loxP/lox5171-flanked T-DNA replacement cassette containing the neomycin phosphotransferase (nptII) coding region via loxP/loxP and lox5171/lox5171 directed recombination. The bar gene is driven by the strong 35S promoter, which is located outside the target cassette. This placement ensures preferential selection of RMCE events and not random integration events by expression of nptII from this same promoter. Using root transformation, during which Cre was provided on a cotransformed T-DNA, 50 kanamycin-resistant calli were selected. Forty-four percent contained a correctly exchanged cassette based on PCR analysis, indicating the stringency of the selection system. This was confirmed for the offspring of five analyzed events by Southern-blot analysis. In four of the five analyzed RMCE events, there were no additional T-DNA insertions or they easily segregated, resulting in high-efficiency single-copy RMCE events. Our approach enables simple and efficient selection of targeting events using the advantages of Agrobacterium-mediated transformation.
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