11. Episomal Expression of Minicircles and Conventional Plasmids in Mammalian Embryos

  1. Dr. Martin Schleef
  1. Wiebke Garrels1,
  2. Khursheed Iqbal2 and
  3. Wilfried A. Kues1

Published Online: 4 APR 2013

DOI: 10.1002/9783527670420.ch11

Minicircle and Miniplasmid DNA Vectors: The Future of Nonviral and Viral Gene Transfer

Minicircle and Miniplasmid DNA Vectors: The Future of Nonviral and Viral Gene Transfer

How to Cite

Garrels, W., Iqbal, K. and Kues, W. A. (2013) Episomal Expression of Minicircles and Conventional Plasmids in Mammalian Embryos, in Minicircle and Miniplasmid DNA Vectors: The Future of Nonviral and Viral Gene Transfer (ed M. Schleef), Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany. doi: 10.1002/9783527670420.ch11

Editor Information

  1. PlasmidFactory GmbH & Co. KG, Dept. R & D, Meisenstr. 96, 33607 Bielefeld, Germany

Author Information

  1. 1

    Friedrich Loeffler Institute, Institute of Farm Animal Genetics, Department of Biotechnology Mariensee, Höltystr. 10, 31535 Neustadt, Germany

  2. 2

    Beckman Research Institute of City of Hope, Arnold and Mabel Beckman Research Center, Department of Molecular and Cellular Biology, Duarte, CA 91010, USA

Publication History

  1. Published Online: 4 APR 2013
  2. Published Print: 17 APR 2013

ISBN Information

Print ISBN: 9783527324569

Online ISBN: 9783527670420

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Keywords:

  • ccc plasmid;
  • cytoplasmic plasmid injection;
  • DNA methylation;
  • epigenetic mechanism;
  • large mammalian model;
  • minicircles;
  • precision genetic engineering;
  • transient transgenesis;
  • transposon

Summary

Genetic engineering of large farm animals is of immense importance for development of improved biomedical models and accelerated breeding progress; however, the available methodological repertoire for genetic modifications is still limited and the molecular events leading to stable transgenesis and sufficient expression are only fragmentarily understood. Currently, pronuclear injection of DNA into zygotes and somatic cell nuclear transfer of genetically modified donor nuclei into enucleated oocytes are standard techniques for the generation of transgenic farm animals. The common principle of both techniques is that they rely on cellular DNA repair mechanisms, which become activated at sites of spontaneous DNA double-stranded breaks. In case of gain-of-function transgenesis, repair-mediated foreign DNA integration might result in silenced or variegated expression patterns. In principle, episomal plasmids and minicircles might be superior for ectopic expression in transgenic animals, because they do not integrate into the chromosomal DNA, thereby avoiding integrational mutagenesis, and are likely to escape silencing mechanisms. This chapter will critically discuss recent developments of nonintegrating episomal vectors for expression in early mammalian embryos.