Short Communication
Molecular Therapy (2006) 13, 625–630; doi: 10.1016/j.ymthe.2005.10.014
RNA as a Source of Transposase for Sleeping Beauty-Mediated Gene Insertion and Expression in Somatic Cells and Tissues
Andrew Wilber1, Joel L. Frandsen1, Jennifer L. Geurts1, David A. Largaespada1, Perry B. Hackett1,2 and R. Scott McIvor1
- 1The Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota, Minneapolis, MN 55455, USA
- 2Discovery Genomics, Inc., Minneapolis, MN 55455, USA
Correspondence: R. Scott McIvor, The Department of Genetics, Cell Biology, and Development, 6-160 Jackson Hall, 321 Church Street SE, University of Minnesota, Minneapolis, MN 55455, USA. Fax: +1 612 625 9810. E-mail: mcivor@mail.med.umn.edu
Received 23 August 2005; Revised 3 October 2005; Accepted 5 October 2005.
Abstract
Sleeping Beauty (SB) is a DNA transposon capable of mediating gene insertion and long-term expression in vertebrate cells when co-delivered with a source of transposase. In all previous reports of SB-mediated gene insertion in somatic cells, the transposase component has been provided by expression of a co-delivered DNA molecule that has the potential for integration into the host cell genome. Integration and continued expression of a gene encoding SB transposase could be problematic if it led to transposon re-mobilization and reintegration. We addressed this potential problem by supplying the transposase-encoding molecule in the form of mRNA. We show that transposase-encoding mRNA can effectively mediate transposition in vitro in HT1080 cells and in vivo in mouse liver following co-delivery with a recoverable transposon or with a luciferase transposon. We conclude that in vitro-transcribed mRNA can be used as an effective source of transposase for SB-mediated transposition in mammalian cells and tissues.
Keywords:
non-viral integration, RNA delivery, bioluminescent imaging, Sleeping Beauty, liver
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