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Stem Cells International
Volume 2011, Article ID 717069, 14 pages
http://dx.doi.org/10.4061/2011/717069
Research Article

Efficient Non-Viral Integration and Stable Gene Expression in Multipotent Adult Progenitor Cells

1Center for Genome Engineering, Gene Therapy Program, Institute of Human Genetics, University of Minnesota, Minneapolis, MN 55455, USA
2Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455, USA
3Department of Medical Microbiology, Immunology and Cell Biology and Simmons Cancer Institute, Southern Illinois University School of Medicine, Springfield, IL 62702, USA
4Department of Medicine, Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455, USA
5Department of Physiology, Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA
6Interdepartmental Stem Cell Institute, Katholieke University Leuven, Leuven, Belgium
7Life Technologies, 5781 Van Allen Way, Carlsbad, CA 92008, USA

Received 14 January 2011; Revised 1 June 2011; Accepted 15 June 2011

Academic Editor: Mark G. Carter

Copyright © 2011 Andrew Wilber et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Non-viral integrating systems, PhiC31 phage integrase ( C31), and Sleeping Beauty transposase (SB), provide an effective method for ex vivo gene delivery into cells. Here, we used a plasmid-encoding GFP and neomycin phosphotransferase along with recognition sequences for both C31 and SB integrating systems to demonstrate that both systems effectively mediated integration in cultured human fibroblasts and in rat multipotent adult progenitor cells (rMAPC). Southern blot analysis of G418-resistant rMAPC clones showed a 2-fold higher number of SB-mediated insertions per clone compared to C31. Sequence identification of chromosomal junction sites indicated a random profile for SB-mediated integrants and a more restricted profile for C31 integrants. Transgenic rMAPC generated with both systems maintained their ability to differentiate into liver and endothelium albeit with marked attenuation of GFP expression. We conclude that both SB and C31 are effective non-viral integrating systems for genetic engineering of MAPC in basic studies of stem cell biology.