Techniques: Recombinogenic engineering–new options for cloning and manipulating DNA

Muyrers, Joep P P; Zhang, Youming; Stewart, A. Francis

doi:10.1016/s0968-0004(00)01757-6

Cited by 229 publications

(160 citation statements)

References 41 publications

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“…To allow rapid assessment of transfection efficiency and selection for stable integration of the mouse BAC in human cells, we designed a universal cassette carrying a neomycin͞kanamycin selection marker and a red fluorescent protein (RFP) marker that carries homologous sequences to replace the chloramphenicol cassette on the BAC backbone by recombineering (14,15). The incorporation of this cassette replaces the internal chloramphenicol resistance gene, thereby allowing a simple selection scheme to obtain successfully modified BACs, monitoring kanamycin resistance and chloramphenicol sensitivity (Fig.…”

Section: Resultsmentioning

confidence: 99%

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RNA interference rescue by bacterial artificial chromosome transgenesis in mammalian tissue culture cells

Kittler

Pelletier

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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RNA interference (RNAi) is a widely used method for analysis of gene function in tissue culture cells. However, to date there has been no reliable method for testing the specificity of any particular RNAi experiment. The ideal experiment is to rescue the phenotype by expression of the target gene in a form refractory to RNAi. The transgene should be expressed at physiological levels and with its different splice variants. Here, we demonstrate that expression of murine bacterial artificial chromosomes in human cells provides a reliable method to create RNAi-resistant transgenes. This strategy should be applicable to all eukaryotes and should therefore be a standard technology for confirming the specificity of RNAi. We show that this technique can be extended to allow the creation of tagged transgenes, expressed at physiological levels, for the further study of gene function.off-target effects ͉ loss-of-function experiment ͉ interferon response ͉ gene knockdown

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Section: Resultsmentioning

confidence: 99%

“…The development of BAC technology for generation of transgenic mice has alleviated this problem to a large extent. Coupled with the development of methodology to specifically mutate large DNA molecules (13), now termed recombineering (14,15), BACs are now the preferred choice to generate transgenic animals (16,17).…”

mentioning

confidence: 99%

RNA interference rescue by bacterial artificial chromosome transgenesis in mammalian tissue culture cells

Kittler

Pelletier

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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“…Because of its simplicity, time-saving capability, and relatively high efficiency, this approach has become a common strategy for introducing mutation into E. coli and C. glutamicum genomes (Ling and Robinson, 1997;Muyrers et al, 2001;Xu et al, 2014b). Although most of the methods used for Single-SDM have been reviewed by researchers around 15-20 years ago (Chatellier et al, 1995;Ling and Robinson, 1997;Muyrers et al, 2001), many new methods are springing up along with the development of genetic engineering technique.…”

Section: Strategies Of Single-sdmmentioning

confidence: 99%

“…In contrast to in vitro genetic modification, it is not limited by the location of the enzyme digestion sites and the construction of base-pairs (Sawitzke et al, 2013). It is executed by introducing recombinant DNA (plasmid or linear DNA) containing selectable marker(s), counter-selectable marker(s), and homologous arm(s) into cells (Datsenko and Wanner, 2000;Yu and Ellis, 2000;Muyrers et al, 2001;Nakashima and Miyazaki, 2014). Recombinant DNA can be constructed in vitro and then introduced into cells.…”

Section: Gene Inactivation Based On Homologous Recombinationmentioning

confidence: 99%

“…Over the past decade, many researchers dedicated themselves to develop the new methods of plasmid-mediated homologous recombination to overcome the defects in the present method (Poustka et al, 1984;Homilton et al, 1989;Kato et al, 1998;Philippe et al, 2004). However, there remain questions on the requirement of long homologous arm(s), how time-consuming it is, low success rates, and the limitation of the recombinogenic window (Muyrers et al, 2001). …”

Section: Plasmid-mediated Homologous Recombinationmentioning

confidence: 99%

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Strategies used for genetically modifying bacterial genome: site-directed mutagenesis, gene inactivation, and gene over-expression

Zhang

2016

J. Zhejiang Univ. Sci. B

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Abstract:With the availability of the whole genome sequence of Escherichia coli or Corynebacterium glutamicum, strategies for directed DNA manipulation have developed rapidly. DNA manipulation plays an important role in understanding the function of genes and in constructing novel engineering bacteria according to requirement. DNA manipulation involves modifying the autologous genes and expressing the heterogenous genes. Two alternative approaches, using electroporation linear DNA or recombinant suicide plasmid, allow a wide variety of DNA manipulation. However, the over-expression of the desired gene is generally executed via plasmid-mediation. The current review summarizes the common strategies used for genetically modifying E. coli and C. glutamicum genomes, and discusses the technical problem of multi-layered DNA manipulation. Strategies for gene over-expression via integrating into genome are proposed. This review is intended to be an accessible introduction to DNA manipulation within the bacterial genome for novices and a source of the latest experimental information for experienced investigators.

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Mouse mutagenesis and gene function

Kühn¹,

Wurst

2005

Encyclopedia of Genetics, Genomics, Proteomics and Bioinformatics

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With the completion of the mouse and human genome sequences, a major challenge is the functional characterization of every gene within the mammalian genome. The mouse offers many advantages for the use of genetics to the study of human biology and disease. A variety of mouse mutagenesis technologies, either gene‐ or phenotype‐driven, are used as systematic approaches. The availability of the mouse genome supports gene‐driven approaches such as gene‐trap and targeted mutagenesis in embryonic stem (ES) cells, allowing an efficiency and precision of gene disruption unmatched among other mammals. Furthermore, chemical and transposon mutagenesis of the mouse genome allows to perform phenotype‐driven screens for the unbiased identification of phenotype–genotype correlations. These technologies already resulted in a collection of several thousand mutants. In this article on mouse mutagenesis and gene function, we present a comprehensive review of gene‐ and phenotype‐driven mouse mutagenesis strategies. Besides a summary of the basic principles of each approach, we emphasize on the latest and future developments of mouse mutagenesis.

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Techniques: Recombinogenic engineering–new options for cloning and manipulating DNA

Cited by 229 publications

References 41 publications

RNA interference rescue by bacterial artificial chromosome transgenesis in mammalian tissue culture cells

RNA interference rescue by bacterial artificial chromosome transgenesis in mammalian tissue culture cells

Strategies used for genetically modifying bacterial genome: site-directed mutagenesis, gene inactivation, and gene over-expression

Mouse mutagenesis and gene function

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