Recombinase-mediated cassette exchange (RMCE) — A rapidly-expanding toolbox for targeted genomic modifications

Turan, Soeren; Zehe, Christoph; Kuehle, Johannes; Jian, Qiao; Bode, Jürgen

doi:10.1016/j.gene.2012.11.016

Cited by 145 publications

(120 citation statements)

References 122 publications

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“…1A) are each capable of undergoing a recombination event with a site of the same kind but not with the other type of site (Schlake and Bode, 1994;Senecoff and Cox, 1986). This type of mutually exclusive recombination between different recombinase target variants has been exploited in several genetic systems, such as iMos (Claveria et al, 2013), Brainbow (Cai et al, 2013;Hadjieconomou et al, 2011;Hampel et al, 2011;Livet et al, 2007), recombinase-mediated cassette exchange (RMCE) (Turan et al, 2013) and the flip-excision (FlEx) transgene inversion strategy (Schnutgen et al, 2005).…”

Section: Design and Features Of The Coinflp Systemmentioning

confidence: 99%

CoinFLP: a system for efficient mosaic screening and for visualizing clonal boundaries in Drosophila

2015

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Section: Design and Features Of The Coinflp Systemmentioning

confidence: 99%

CoinFLP: a system for efficient mosaic screening and for visualizing clonal boundaries in Drosophila

2015

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“…The creation of a transgenic mouse line can be achieved by a wide spectrum of technologies including random integration of a transgene via pronuclear injection, a more precise and predictable homologous recombination, or, as established in the last decade, recombinase-mediated cassette exchange (RMCE) (21), by using zinc-finger nuclease (ZFN), Transcription Activator-like Effector Nuclease (TALEN) and most recently CRISPR/Cas technologies (22).…”

Section: Transgenic Animalsmentioning

confidence: 99%

“…It can be achieved by precisely replacing a genomic target cassette by a compatible donor construct (21).…”

Section: Recombinase-mediated Cassette Exchangementioning

confidence: 99%

Suppression of hepatic fibrosis by efficient Col1a1 silencing using shRNA inducible mouse models

Molokanova¹,

Shi²,

Weng³

et al. 2017

Journal of Hepatology

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“…The Cre/loxP system for site-specific DNA recombination has been known for over two decades as a genome modification tool for animal cells (Turan et al, 2013). We have developed a retroviral vector that targets DNA into a pre-determined site of engineered target cells using a combination of an integrase-defective retroviral vector (IDRV) and Cre recombinase-mediated cassette exchange (RMCE) (Huang et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Targeted transgene insertion into the CHO cell genome using Cre recombinase‐incorporating integrase‐defective retroviral vectors

Kawabe

Shimomura

Huang

et al. 2016

Biotech & Bioengineering

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Retroviral vectors have served as efficient gene delivery tools in various biotechnology fields. However, viral DNA is randomly inserted into the genome, which can cause problems, such as insertional mutagenesis and gene silencing. Previously, we reported a site-specific gene integration system, in which a transgene is integrated into a predetermined chromosomal locus of Chinese hamster ovary (CHO) cells using integrase-defective retroviral vectors (IDRVs) and Cre recombinase. In this system, a Cre expression plasmid is transfected into founder cells before retroviral transduction. In practical applications of site-specific gene modification such as for hard-to-transfect cells or for in vivo gene delivery, both the transgene and the Cre protein into retroviral virions should be encapsulate. Here, we generated novel hybrid IDRVs in which viral genome and enzymatically active Cre can be delivered (Cre-IDRVs). Cre-IDRVs encoding marker genes, neomycin resistance and enhanced green fluorescent protein (EGFP), flanked by wild-type and mutated loxP sites were produced using an expression plasmid for a chimeric protein of Cre and retroviral gag-pol. After analyzing the incorporation of the Cre protein into retroviral virions by Western blotting, the Cre-IDRV was infected into founder CHO cells, in which marker genes (hygromycin resistance and red fluorescent protein) flanked with corresponding loxP sites are introduced into the genome. G418-resistant colonies expressing GFP appeared and the site-specific integration of the transgene into the expected chromosomal site was confirmed by PCR and sequencing of amplicons. Moreover, when Cre-IDRV carried a gene expression unit for a recombinant antibody, the recombinant cells in which the antibody expression cassette was integrated in a site-specific manner were generated and the cells produced the recombinant antibody. This method may provide a promising tool to perform site-specific gene modification according to Cre-based cell engineering. Biotechnol. Bioeng. 2016;113: 1600-1610. © 2016 Wiley Periodicals, Inc.

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Recombinase-mediated cassette exchange (RMCE) — A rapidly-expanding toolbox for targeted genomic modifications

Cited by 145 publications

References 122 publications

CoinFLP: a system for efficient mosaic screening and for visualizing clonal boundaries in Drosophila

CoinFLP: a system for efficient mosaic screening and for visualizing clonal boundaries in Drosophila

Suppression of hepatic fibrosis by efficient Col1a1 silencing using shRNA inducible mouse models

Targeted transgene insertion into the CHO cell genome using Cre recombinase‐incorporating integrase‐defective retroviral vectors

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