Coping with kinetic and thermodynamic barriers: RMCE, an efficient strategy for the targeted integration of transgenes

Baer, Alexandra; Bode, Jürgen

doi:10.1016/s0958-1669(00)00248-2

Cited by 138 publications

(112 citation statements)

References 41 publications

Supporting

Mentioning

110

Contrasting

Unclassified

Order By: Relevance

“…An efficient gene targeting method for insects was based on a recombinase-mediated cassette exchange (RMCE) strategy first developed for genomic targeting in mammalian stem cells (Baer and Bode 2001). We first tested RMCE in Drosophila (Horn and Handler 2005) using the FRT/FLP recombination system (Andrews et al 1985;Siegal and Hartl 1996) by designing a transposon acceptor vector that once integrated by piggyBac transformation (Handler et al 1998), could act as a target-site for subsequent donor vector insertions by double-recombination of variant FRT sites (see Horn and Handler 2005 , Fig.…”

Section: Rmce Targeting and Stabilization Vectorsmentioning

confidence: 99%

Not all GMOs are crop plants: non-plant GMO applications in agriculture

et al. 2013

View full text Add to dashboard Cite

Since tools of modern biotechnology have become available, the most commonly applied and often discussed genetically modified organisms are genetically modified crop plants, although genetic engineering is also being used successfully in organisms other than plants, including bacteria, fungi, insects, and viruses. Many of these organisms, as with crop plants, are being engineered for applications in agriculture, to control plant insect pests or diseases. This paper reviews the genetically modified non-plant organisms that have been the subject of permit approvals for environmental release by the United States Department of Agriculture/Animal and Plant Health Inspection Service since the US began regulating genetically modified organisms. This is an indication of the breadth and progress of research in the area of non-plant genetically modified organisms. This review includes three examples of promising research on non-plant genetically modified organisms for application in agriculture: (1) insects for insect pest control using improved vector systems; (2) fungal pathogens of insects to control insect pests; and (3) virus for use as transient-expression vectors for disease control in plants

show abstract

Section: Rmce Targeting and Stabilization Vectorsmentioning

confidence: 99%

Not all GMOs are crop plants: non-plant GMO applications in agriculture

et al. 2013

View full text Add to dashboard Cite

show abstract

“…However, the use of two inverted loxP sites or a pair of mutant loxPs can increase the integration efficiency of target DNA. This system, called RMCE, enables exchange of a preexisting DNA cassette flanked by mutant loxPs with the DOI flanked by compatible loxPs [7,76]. Since the integration efficiency in RMCE has been proven to be much higher than that in homologous recombination-based targeting, targeting of DOis with RMCE is now frequently performed using ES cells containing loxP-tagged sites.…”

Section: Cre-loxp-based Pitt (Cre-pitt)mentioning

confidence: 99%

PITT: Pronuclear Injection-Based Targeted Transgenesis, a Reliable Transgene Expression Method in Mice

et al. 2012

View full text Add to dashboard Cite

Transgenic (Tg) mice have been extensively used as valuable tools for analyses of gene function and have also served as models for many human diseases. Typically, a transgenic mouse is created by microinjection of DNA into pronuclei in which the DNA gets integrated at random locations in the genome. Frequently however, the random integration of multiple copies of a transgene results in transgene silencing, probably because of a positional effect and/or repeat-induced gene silencing. The transgene silencing issue has been overcome by single-copy transgene integration into a predetermined locus through ES cell-mediated transgenesis, despite it being expensive and more time-consuming compared with pronuclear injection (PI)-mediated transgenesis. Recently, several groups have reported novel approaches that employ PI for targeted transgenesis. They are based on site-specific recombination catalyzed by a recombinase or an integrase or homologous recombination enhanced by a zinc-finger nuclease via PI. These next-generation transgenesis methods, which we termed as PI-based Targeted Transgenesis (PITT), are more convenient and faster than ES cell-based transgenesis. Furthermore, the Tg mice generated by these newer methods contain a single-copy transgene and exhibit reliable expression of the transgene. The objective of this review is to present the recent progress in mouse targeted transgenesis.

show abstract

“…Thus the copy number of an integrated gene may not indicate a stable expression since deterioration over time may occur due to repeat induced silencing. Targeting transgene integration by the use of the Cre/loxP recombination system and Flp/FRT (Flp-recognition target) in combination with recombinase mediated cassette exchange (RCME) (Baer and Bode 2001) means it is possible to create cell lines which can be selected for high expression of a specific tag, such as Green Fluorescent Protein (GFP). The selected clones can be assessed for stability and are transfected with the gene of interest so that RCME takes place and a high producing cell line with the gene of interest is obtained (Baer andBode 2001, Feng et al 1999).…”

Section: Expression Engineeringmentioning

confidence: 99%

Using cell engineering and omic tools for the improvement of cell culture processes

et al. 2007

View full text Add to dashboard Cite

Significant strides have been made in mammalian cell based biopharmaceutical process and cell line development over the past years. With several established mammalian host cell lines and expression systems, optimization of selection systems to reduce development times and improvement of glycosylation patterns are only some of the advances being made to improve cell culture processes. In this article, the advances pertaining to cell line development and cell engineering strategies are discussed. An overview of the cell engineering strategies to enhance cellular characteristics by genetic manipulation are illustrated, focusing on the use of genomics and proteomics tools and their application in such endeavors. Included in this review are some of the early studies using the 'omic' technique to understand cellular mechanisms of product synthesis and secretion, apoptosis, cell proliferation and the influence of the physicochemical environment. The article highlights the significance of integrating genomics and proteomics data with the vast amounts of bioprocess data for improved analysis of the biological pathways involved. Further improvements of the techniques and methodologies used are needed but ultimately, the new cell engineering strategies should provide great insight into the regulatory networks within the cell in a bioprocess environment and how to manipulate them to increase overall productivity.

show abstract

Coping with kinetic and thermodynamic barriers: RMCE, an efficient strategy for the targeted integration of transgenes

Cited by 138 publications

References 41 publications

Not all GMOs are crop plants: non-plant GMO applications in agriculture

Not all GMOs are crop plants: non-plant GMO applications in agriculture

PITT: Pronuclear Injection-Based Targeted Transgenesis, a Reliable Transgene Expression Method in Mice

Using cell engineering and omic tools for the improvement of cell culture processes

Contact Info

Product

Resources

About