ArticleNucleic acids research2012
Flp and Cre expressed from Flp-2A-Cre and Flp-IRES-Cre transcription units mediate the highest level of dual recombinase-mediated cassette exchange.
Article in Nucleic acids research, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
27 citing papers in PubMed.
- Cell Fate Determination and Lineage Tracing: Technological Evolution and Multidimensional Applications.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Review
- Recent advances in therapeutic gene-editing technologies.Molecular therapy : the journal of the American Society of Gene Therapy · 2025Review
- A powerful and highly efficient PAI-mediated transgenesis approach in Drosophila.Nucleic acids research · 2025Article
- Large T antigen mediated target gene replication improves site-specific recombination efficiency.Frontiers in bioengineering and biotechnology · 2024Article
- A cell competition system with one gene expression from a single-copy gene in one cell.PloS one · 2024Article
- High-fidelity large-diversity monoclonal mammalian cell libraries by cell cycle arrested recombinase-mediated cassette exchange.Nucleic acids research · 2023Article
- Discovery and characterization of novel Cre-type tyrosine site-specific recombinases for advanced genome engineering.Nucleic acids research · 2023Article
- Systematic discovery of recombinases for efficient integration of large DNA sequences into the human genome.Nature biotechnology · 2023Article
- Article
- Pluripotent stem cell-based gene therapy approach: human de novo synthesized chromosomes.Cellular and molecular life sciences : CMLS · 2021Review
- Article
- Genome targeting by hybrid Flp-TAL recombinases.Scientific reports · 2020Article
- Article
- Single Copy Transgene Integration in a Transcriptionally Active Site for Recombinant Protein Synthesis.Biotechnology journal · 2018Article
- Flexible pseudotyping of retrovirus using recombinase-mediated cassette exchange.Biotechnology letters · 2018Article
- Improving the efficiency of gene insertion in a human artificial chromosome vector and its transfer in human-induced pluripotent stem cells.Biology methods & protocols · 2018Article
- Efficient Genome Manipulation by Variants of Site-Specific Recombinases R and TD.Journal of molecular biology · 2016Article
- Bone morphogenetic protein 9 (BMP9) induces effective bone formation from reversibly immortalized multipotent adipose-derived (iMAD) mesenchymal stem cells.American journal of translational research · 2016Article
- Redesign of the monomer-monomer interface of Cre recombinase yields an obligate heterotetrameric complex.Nucleic acids research · 2015Article
- Target-specific variants of Flp recombinase mediate genome engineering reactions in mammalian cells.The FEBS journal · 2015Article
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3 authors.
Funding
Abstract
Recombinase-mediated cassette exchange (RMCE) is a powerful tool for unidirectional integration of DNA fragments of interest into a pre-determined genome locale. In this report, we examined how the efficiency of dual RMCE catalyzed by Flp and Cre depends on the nature of transcription units that express the recombinases. The following recombinase transcription units were analyzed: (i) Flp and Cre genes expressed as individual transcription units located on different vectors, (ii) Flp and Cre genes expressed as individual transcription units located on the same vector, (iii) Flp and Cre genes expressed from a single promoter and separated by internal ribosome entry sequence and (iv) Flp and Cre coding sequences separated by the 2A peptide and expressed as a single gene. We found that the highest level of dual RMCE (35-45% of the transfected cells) can be achieved when Flp and Cre recombinases are expressed as Flp-2A-Cre and Flp-IRES-Cre transcription units. In contrast, the lowest level of dual RMCE (∼1% of the transfected cells) is achieved when Flp and Cre are expressed as individual transcription units. The analysis shows that it is the relative Flp-to-Cre ratio that critically affects the efficiency of dual RMCE. Our results will be helpful for maximizing the efficiency of dual RMCE aimed to engineer and re-engineer genomes.
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