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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board Member: Buchner, Johannes / Ludwig, Stephan / Sies, Helmut / Turk, Boris / Wittinghofer, Alfred

12 Issues per year

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The Transgeneticists Toolbox: Novel Methods for the Targeted Modification of Eukaryotic Genomes

Jürgen Bode / Thomas Schlake / Michaela Iber / Dirk Schübeler / Jost Seibler / Evgeney Snezhkov / Lev Nikolaev

Citation Information: Biological Chemistry. Volume 381, Issue 9-10, Pages 801–813, ISSN (Print) 1431-6730, DOI: 10.1515/BC.2000.103, July 2005

Publication History

Published Online:
2005-07-05

Abstract

Classical techniques for gene transfer into mammalian cells involve tedious screening procedures to identify transgenic clones or animals with the appropriate level and stability of expression or with the correct developmental patterns. These first generation technologies are clearly inadequate for complex genetic strategies by which gene regulation can be studied in its entire complexity. While site-specific insertions can principally be achieved by homologous recombination or by adapting the recombination apparatus from phages or yeast, these methods usually lack the required efficiency or they perturb expression patterns by the co-insertion of prokaryotic vector parts. Virtually all of these problems can be overcome by recombinase-mediated cassette exchange (RMCE) techniques which cleanly replace a resident cassette that is flanked by two hetero-specific recombination target sites for a second cassette with the analogous design, presented on a targeting vector. After illustrating the fundamentals of site-specific recombination by selected experiments, the authors (arranged in the chronological order of their contribution) will describe their efforts to develop RMCE into a method of wide applicability. Further developments that have been initiated utilizing the particular potential of the RMCE principle will be outlined.

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